The following description of the background of the invention is provided to aid in understanding the invention, but is not admitted to describe or constitute prior art to the invention.
Protein kinases (xe2x80x9cPKsxe2x80x9d) are enzymes that catalyze the phosphorylation of hydroxy groups on tyrosine, serine and threonine residues of proteins. The consequences of this seemingly simple activity are staggering. Cell growth, differentiation and proliferation, i.e., virtually all aspects of cell life, in one way or another depend on PK activity. Furthermore, abnormal PK activity has been related to a host of disorders, ranging from relatively non-life-threatening diseases such as psoriasis to extremely virulent diseases such as glioblastoma (brain cancer).
The PKs can conveniently be broken down into two classes, the protein tyrosine kinases (PTKs) and the serine-threonine kinases (STKs).
One of the prime aspects of PK activity is involvement with growth factor receptors. Growth factor receptors are cell-surface proteins. When bound by a growth factor ligand, growth factor receptors are converted to an active form that can interact with proteins on the inner surface of a cell membrane. This interaction leads to phosphorylation on tyrosine residues of the receptor as well as other amino acids and to the formation inside the cell of complexes with a variety of cytoplasmic signaling molecules. In turn, these complexes affect numerous cellular responses such as cell division (proliferation), cell differentiation, cell growth, expression of metabolic effects on the extracellular microenvironment, etc. For a more complete discussion, see Schlessinger and Ullrich, Neuron, 1992, 9:303-391 which is incorporated by reference, including any drawings, as if fully set forth herein.
Receptor tyrosine kinases (RTKs) are growth factor receptors with PK activity. They comprise a large family of transmembrane receptors with diverse biological activity. At present, at least nineteen (19) distinct subfamilies of RTKs have been identified. An example of these is the subfamily designated the xe2x80x9cHERxe2x80x9d RTKs, which includes EGFR (epithelial growth factor receptor), HER2, HER3 and HER4. These RTKs consist of an extracellular glycosylated ligand binding domain, a transmembrane domain and an intracellular cytoplasmic catalytic domain that can phosphorylate tyrosine residues on proteins.
Another RTK subfamily consists of insulin receptor (IR), insulin-like growth factor I receptor (IGF-1R) and insulin receptor related receptor (IRR). IR and IGF-1R interact with insulin, IGF-I and IGF-II to form a heterotetramer composed of two entirely extracellular glycosylated xcex1 subunits and two xcex2 subunits which contain the tyrosine kinase domain.
A third RTK subfamily is referred to as the platelet derived growth factor receptor (xe2x80x9cPDGFRxe2x80x9d) group, which includes PDGFRxcex1, PDGFRxcex2, CSFIR, c-kit and c-fms. These receptors consist of a glycosylated extracellular domain composed of variable numbers of immunoglobin-like loops, a transmembrane domain and an intracellular domain having a tyrosine kinase domain interrupted by unrelated amino acid sequences.
Another group which, because of its similarity to the PDGFR subfamily, is sometimes subsumed in the latter group, is the fetus liver kinase (xe2x80x9cflkxe2x80x9d) receptor subfamily. This group is believed to be composed of kinase insert domain-receptor fetal liver kinase-1 (KDR/FLK-1), flk-1R, flk-4 and fms-like tyrosine kinase 1 (flt-1).
One further member of the tyrosine kinase growth factor receptor family is the fibroblast growth factor (xe2x80x9cFGFxe2x80x9d)receptor group. This group consists of four receptors, FGFR1-4, and seven ligands, FGF1-7. While not yet well characterized, it appears that the receptors also consist of a glycosylated extracellular domain containing a variable number of immunoglobin-like loops, a transmembrane domain and an intracellular domain in which the tyrosine kinase domain is interrupted by regions of unrelated amino acid sequences.
A more complete listing of the known RTK subfamilies is described in Plowman et al., DNandP, 1994, 7(6):334-339 which is incorporated by reference, including any drawings, as if fully set forth herein.
In addition to the RTKs, there also exists a family of entirely intracellular PTKs called xe2x80x9cnon-receptor tyrosine kinasesxe2x80x9d or xe2x80x9ccellular tyrosine kinasesxe2x80x9d (xe2x80x9cCTKxe2x80x9d). CTKs do not contain extracellular and transmembrane domains. At present, over 24 CTKs in 11 subfamilies (Src, Frk, Btk, Csk, Abl, Zap70, Fes, Fps, Fak, Jak and Ack) have been identified. The Src subfamily appears so far to be the largest group of CTKs and includes Src, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk. For a more detailed discussion of CTKs, see Bolen, Oncogene, 1993, 8:2025-2031, which is incorporated by reference, including any drawings, as if fully set forth herein.
The serine-threonine kinases or STKs, like the CTKs, are predominantly intracellular although there are a few STK receptor kinases. STKs are the most common of the cytosolic kinases; i.e., kinases that perform their function in that part of the cytoplasm other than the cytoplasmic organelles and cytoskelton. The cytosol is the region within the cell where much of the cell""s intermediary metabolic and biosynthetic activity occurs; e.g., it is in the cytosol that proteins are synthesized on ribosomes.
RTKs, CTKs and STKs have all been implicated in a host of pathogenic conditions including, significantly, cancer. Other pathogenic conditions which have been associated with PTKs include, without limitation, psoriasis, hepatic cirrhosis, diabetes, atherosclerosis, angiogenesis, restenosis, ocular diseases, rheumatoid arthritis and other inflammatory disorders, immunological disorders such as autoimmune disease, cardiovascular diseases such as atherosclerosis and a variety of renal disorders.
With regard to cancer, two of the major hypotheses advanced to explain the excessive cellular proliferation that drives tumor development relate to functions known to be PK regulated. That is, it has been suggested that malignant cell growth is the result of a breakdown in the mechanisms that control cell division and/or differentiation. It has been shown that the protein products of a number of proto-oncogenes are involved in the signal transduction pathways that regulate cell growth and differentiation. These protein products of proto-oncogenes include the extracellular growth factors, transmembrane growth factor PTK receptors (RTKs), cytoplasmic PTKs (CTKs) and cytosolic STKs, discussed above.
In view of the apparent link between PK-related cellular activities and a number of human disorders, it is no surprise that a great deal of effort is being spent to identify ways to modulate PK activity. Some of these efforts have been directed at biomimetic approaches using large molecules patterned on those involved in the actual cellular processes (e.g., mutant ligands (U.S. Pat. No. 4,966,849); soluble receptors and antibodies (App. No. WO 94/10202, Kendall and Thomas, Proc. Nat""l Acad. Sci., 1994, 90:10705-09, Kim, et al., Nature, 1993, 362:841-844); RNA ligands (Jelinek, et al., Biochemistry, 33:10450-56); Takano, et al., Mol. Bio. Cell, 1993, 4:358A; Kinsella, et al., Exp. Cell Res., 1992, 199:56-62; Wright, et al., J. Cellular Phys., 152:448-57) and tyrosine kinase inhibitors (WO 94/03427; WO 92/21660; WO 91/15495; WO 94/14808; U.S. Pat. No. 5,330,992; Mariani, et al., Proc. Am. Assoc. Cancer Res., 1994, 35:2268).
More recently, attempts have been made to identify small molecules that act as PK inhibitors. For example, bis-monocylic, bicyclic and heterocyclic aryl compounds (PCT WO 92/20642), vinylene-azaindole derivatives (PCT WO 94/14808) and 1-cyclopropyl-4-pyridylquinolones (U.S. Pat. No. 5,330,992) have been described as PTK inhibitors. Styryl compounds (U.S. Pat. No. 5,217,999), styryl-substituted pyridyl compounds (U.S. Pat. No. 5,302,606), quinazoline derivatives (EP App. No. 0 566 266 A1), selenaindoles and selenides (PCT WO 94/03427), tricyclic polyhydroxylic compounds (PCT WO 92/21660) and benzylphosphonic acid compounds (PCT WO 91/15495) have all been described as PTK inhibitors useful in the treatment of cancer.
The present invention is directed in part towards indolinone compounds and methods of modulating the function of protein kinases with these compounds. The methods incorporate cells that express a protein kinase. In addition, the invention describes methods of preventing and treating protein kinases-related abnormal conditions in organisms with a compound identified by the methods described herein. Furthermore, the invention pertains to pharmaceutical compositions comprising compounds identified by methods of the invention.
The present invention features indolinone compounds that potently inhibit protein kinases and related products and methods. Inhibitors of protein kinases can be obtained by adding chemical substituents to an indolinone compound. The compounds of the invention represent a new generation of therapeutics for diseases associated with one or more functional or non-functional protein kinases. Neuro-degenerative diseases and certain types of cancer fall into this class of diseases. Other diseases or disorders include dermatologic, ophthalmic, nurologic, cardiovascular, and immune disorders as well as disorders associated with abnormal angiogenesis and/or vasculogenesis. The compounds can be modified such that they are specific to their target or targets and will subsequently cause few side effects and thus represent a new generation of potential cancer therapeutics. These properties are significant improvements over the currently utilized cancer therapeutics that cause multiple side effects and deleteriously weaken patients.
It is believed the compounds of the invention will minimize or obliterate solid tumors by inhibiting the activity of the protein kinases, or will at least modulate or inhibit tumor growth and/or metastases. Protein kinases regulate proliferation of blood vessels during angiogenesis, among other functions. Increased rates of angiogenesis accompany cancer tumor growth in cells as cancer tumors must be nourished by oxygenated blood during growth. Therefore, inhibition of the protein kinase and the corresponding decreases in angiogenesis will starve tumors of nutrients and most likely obliterate them.
While a precise understanding of the mechanism by which compounds inhibit PTKs (e.g., the fibroblast growth factor receptor 1 [FGFR 1]) is not required in order to practice the present invention, the compounds are believed to interact with the amino acids of the PTKs"" catalytic region. PTKs typically possess a bi-lobate structure, and ATP appears to bind in the cleft between the two lobes in a region where the amino acids are conserved among PTKs; inhibitors of PTKs are believed to bind to the PTKs through non-covalent interactions such as hydrogen bonding, Van der Waals interactions, and ionic bonding, in the same general region that ATP binds to the PTKs. More specifically, it is thought that the oxindole component of the compounds of the present invention binds in the same general space occupied by the adenine ring of ATP. Specificity of a PTK inhibitor for a particular PTK may be conferred by interactions between the constituents around the oxindole core with amino acid domains specific to individual PKs. Thus, different substitutents may contribute to preferential binding to particular PKs. The ability to select those compounds active at different ATP binding sites makes them useful in targeting any protein with such a site, including not only protein tyrosine kinases, but also serine/threonine kinases. Thus, such compounds have utility for in vitro assays on such proteins and for in vivo therapeutic effect through such proteins.
Usually, indolinone compounds that are synthesized by the condensation of an oxindole compound and a ketone compound display a mixture of the possible E and Z isomers, making the isolation of the isomer of choice difficult. The compounds of the present invention feature an intermolecular hydrogen bond between the carbonyl of the oxindole compound and the hydrogen of the 1 position of the pyrrole moiety of the ketone compounds. Said hydrogen bond eliminates the problem of having a mixture of isomers by locking the intermediates in the synthesis of the indolinone compounds in the preferred conformation.
The term xe2x80x9ccompoundxe2x80x9d refers to the compound or a pharmaceutically acceptable salt, ester, amide, prodrug, isomer, or metabolite, thereof.
The term xe2x80x9cpharmaceutically acceptable saltxe2x80x9d refers to a formulation of a compound that does not abrogate the biological activity and properties of the compound. Pharmaceutical salts can be obtained by reacting a compound of the invention with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.
A xe2x80x9cprodrugxe2x80x9d refers to an agent that is converted into the parent drug in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. An example, without limitation, of a prodrug would be a compound of the present invention which is administered as an ester (the xe2x80x9cprodrugxe2x80x9d) to facilitate transmittal across a cell membrane where water solubility is detrimental to mobility but which then is metabolically hydrolyzed to the carboxylic acid, the active entity, once inside the cell where water solubility is beneficial. A fruther example of a prodrug might be a short peptide (polyaminoacid) bonded to an acid group wherein the peptide is metabolized to reveal the active moiety.
The term xe2x80x9cindolinonexe2x80x9d is used as that term is commonly understood in the art and includes a large subclass of substituted or unsubstituted compounds that are capable of being synthesized from an aldehyde moiety and an oxindole moiety.
The term xe2x80x9coxindolexe2x80x9d refers to an oxindole compound substituted with chemical substituents. Oxindole compounds are of the general structure: 
As used herein a xe2x80x9cfused pyrroloxe2x80x9d group refers to the structure in brackets: 
The term xe2x80x9csubstitutedxe2x80x9d, in reference to the invention, refers to an oxindole compound that is derivatized with any number of chemical substituents.
As used herein, the term xe2x80x9calkylxe2x80x9d refers to an aliphatic hydrocarbon group. The alkyl moiety may be a xe2x80x9csaturated alkylxe2x80x9d group, which means that it does not contain any alkene or alkyne moieties. The alkyl moiety may also an be xe2x80x9cunsaturated alkylxe2x80x9d moiety, which means that it contains at least one alkene or alkyne moiety. An xe2x80x9calkenexe2x80x9d moiety refers to a group consisting of at least two carbon atoms and at least one carbon-carbon double bond, and an xe2x80x9calkynexe2x80x9d moiety refers to a groupconsisting of at least two carbon atoms and at least one carbon-carbon triple bond. The alkyl moiety, whether saturated or unsaturated, may be branched, non-branched, or cyclic.
Preferably, the alkyl group has 1 to 20 carbon atoms (whenever it appears herein, a numerical range such as xe2x80x9c1 to 20xe2x80x9d refers to each integer in the given range; e.g., xe2x80x9c1 to 20 carbon atomsxe2x80x9d means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms, although the present definition also covers the occurrence of the term xe2x80x9calkylxe2x80x9d where no numerical range is designated). More preferably, it is a medium size alkyl having 1 to 10 carbon atoms. Most preferably, it is a lower alkyl having 1 to 4 carbon atoms. The alkyl group may be substituted or unsubstituted. When substituted, the substituent group(s) is(are) preferably one or more group(s) individually and independently selected from cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, nitro, silyl and amino, including mono- and di-substituted amino groups.
The term xe2x80x9caromaticxe2x80x9d refers to an aromatic group which has at least one ring having a conjugated pi electron system and includes both carbocyclic aryl (e.g., phenyl) and heterocyclic aryl groups (e.g., pyridine). The term includes monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) groups. The term xe2x80x9ccarbocyclicxe2x80x9d refers to a compound which contains one or more covalently closed ring structures, and that the atoms forming the backbone of the ring are all carbon atoms. The term thus distinguishes carbocyclic from heterocyclic rings in which the ring backbone contains at least one atom which is different from carbon. The term xe2x80x9cheteroaromaticxe2x80x9d refers to an aromatic group which contains at least one heterocyclic ring.
The term xe2x80x9caliphatic ringxe2x80x9d refers to a compound which contains one or more covalently closed ring structures, and that at least one of the atoms forming the backbone is a saturated carbon atom (e.g., cyclohexane). The term xe2x80x9cheteroaliphatic ringxe2x80x9d refers to a ring system in which at least one of the atoms forming the backbone is a heteroatom (e.g., tetrahydropyran).
The term xe2x80x9caminexe2x80x9d refers to a chemical moiety of formula xe2x80x94NR1R2 where R1 and R2 are independently selected from the group consisting of hydrogen, saturated or unsaturated alkyl, and five-membered or six-membered aryl or heteroaryl ring moieties, where the ring is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, nitro, and ester moieties.
The term xe2x80x9ciminexe2x80x9d refers to a chemical moiety of formula xe2x80x94Nxe2x95x90R1 where R1 is selected from the group consisting of hydrogen, saturated or unsaturated alkyl, and aryl or heteroaryl ring moieties (monocyclic or bicyclic), where the ring is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, nitro, and ester moieties.
The term xe2x80x9chalogenxe2x80x9d or xe2x80x9chaloxe2x80x9d refers to an atom selected from the group consisting of fluorine, chlorine, bromine, and iodine. The term xe2x80x9ctrihalomethylxe2x80x9d refers to the xe2x80x94CX3 group, where X is a halogen.
The term xe2x80x9ccarboxylic acidxe2x80x9d refers to a chemical moiety with formula xe2x80x94(R)nxe2x80x94COOH, where R is selected from the group consisting of of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) and heteroalicyclic (bonded through a ring carbon), as those terms are defined herein, and where n is 0 or 1.
The term xe2x80x9cesterxe2x80x9d refers to a chemical moiety with formula xe2x80x94(R)nxe2x80x94COORxe2x80x2, where R and Rxe2x80x2 are independently selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) and heteroalicyclic (bonded through a ring carbon), as those terms are defined herein, and where n is 0 or 1. As used herein, the term xe2x80x9ccarboxyalkylxe2x80x9d also falls within this definition.
The term xe2x80x9caldehydexe2x80x9d refers to a chemical moiety with formula xe2x80x94(R)nxe2x80x94CHO, where R is as defined herein and where n is 0 or 1.
The term xe2x80x9csulfonexe2x80x9d refers to a chemical moiety with formula xe2x80x94SO2xe2x80x94R, where R is as defined herein.
The term xe2x80x9cacylxe2x80x9d refers to chemical moieties of the general formula xe2x80x94C(O)R. When R is hydrogen the molecule containing the acyl group is an aldehyde. When R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) and heteroalicyclic (bonded through a ring carbon), as those terms are defined herein, then the molecule containing the acyl group is a ketone.
A xe2x80x9cthiocarbonylxe2x80x9d group refers to a xe2x80x94C(xe2x95x90S)xe2x80x94R group, where R is as defined herein.
An xe2x80x9cO-carboxyxe2x80x9d group refers to a RC(xe2x95x90O)Oxe2x80x94 group, where R is as defined herein.
A xe2x80x9cC-carboxyxe2x80x9d group refers to a xe2x80x94C(xe2x95x90O)OR groups where R is as defined herein.
An xe2x80x9cacetylxe2x80x9d group refers to a xe2x80x94C(xe2x95x90O)CH3 group.
A xe2x80x9ctrihalomethanesulfonylxe2x80x9d group refers to a X3CS(xe2x95x90O)2xe2x80x94 group where X is a halogen.
A xe2x80x9ccyanoxe2x80x9d group refers to a xe2x80x94Cxe2x89xa1N group.
An xe2x80x9cisocyanatoxe2x80x9d group refers to a xe2x80x94NCO group.
A xe2x80x9cthiocyanatoxe2x80x9d group refers to a xe2x80x94CNS group.
An xe2x80x9cisothiocyanatoxe2x80x9d group refers to a xe2x80x94NCS group.
A xe2x80x9csulfinylxe2x80x9d group refers to a xe2x80x94S(xe2x95x90O)xe2x80x94R group, with R as defined herein.
A xe2x80x9cS-sulfonamidoxe2x80x9d group refers to a xe2x80x94S(xe2x95x90O)2NR2 group, with R as defined herein.
A xe2x80x9cN-sulfonamidoxe2x80x9d group refers to a RS(xe2x95x90O)2NHxe2x80x94 group with R as defined herein.
A xe2x80x9ctrihalomethanesulfonamidoxe2x80x9d group refers to a X3CS(xe2x95x90O)2NRxe2x80x94 group with X and R as defined herein.
An xe2x80x9cO-carbamylxe2x80x9d group refers to a xe2x80x94OC(xe2x95x90O)xe2x80x94NR2 group with R as defined herein.
An xe2x80x9cN-carbamylxe2x80x9d group refers to a ROC(xe2x95x90O)NHxe2x80x94 group, with R as defined herein.
An xe2x80x9cO-thiocarbamylxe2x80x9d group refers to a xe2x80x94OC(xe2x95x90S)xe2x80x94NR2 group with R as defined herein.
An xe2x80x9cN-thiocarbamylxe2x80x9d group refers to an ROC(xe2x95x90S)NHxe2x80x94 group, with R as defined herein.
A xe2x80x9cC-amidoxe2x80x9d group refers to a xe2x80x94C(xe2x95x90O)xe2x80x94NR2 group with R as defined herein.
An xe2x80x9cN-amidoxe2x80x9d group refers to a RC(xe2x95x90O)NHxe2x80x94 group, with R as defined herein.
By xe2x80x9ccombined,xe2x80x9d when referring to two adjacent xe2x80x9cRxe2x80x9d groups herein is meant that the two xe2x80x9cRxe2x80x9d groups are covalently bonded to each other so as to form a ring system. The ring system may be cycloalkyl, aryl, heteroaryl or heteroalicyclic.
A xe2x80x9ccombinatorial libraryxe2x80x9d refers to all the compounds formed by the reaction of each compound of one dimension with a compound in each of the other dimensions in a multi-dimensional array of compounds. In the context of the present invention, the array is two dimensional and one dimension represents all the oxindoles of the invention and the second dimension represents all the aldehydes of the invention. Each oxindole may be reacted with each and every aldehyde in order to form an indolinone compound. All indolinone compounds formed in this way are within the scope of the present invention. Also within the scope of the present invention are smaller combinatorial libraries formed by the reaction of some of the oxindoles with all of the aldehydes, all of the oxindoles with some of the aldehydes, or some of the oxindoles with some of the aldehydes.
The term xe2x80x9cpharmaceutical compositionxe2x80x9d refers to a mixture of a compound of the invention with other chemical components, such as diluents, excipients, or carriers. The pharmaceutical composition facilitates administration of the compound to an organism. Multiple techniques of administering a compound exist in the art including, but not limited to, oral, aerosol, parenteral, and topical administration. Pharmaceutical compositions can also be obtained by reacting compounds with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.
The term xe2x80x9cphysiologically acceptablexe2x80x9d defines a carrier or diluent that does not abrogate the biological activity and properties of the compound.
The term xe2x80x9ccarrierxe2x80x9d defines a chemical compound that facilitates the incorporation of a compound into cells or tissues. For example dimethyl sulfoxide (DMSO) is a commonly utilized carrier as it facilitates the uptake of many organic compounds into the cells or tissues of an organism.
The term xe2x80x9cdiluentxe2x80x9d defines chemical compounds diluted in water that will dissolve the compound of interest as well as stabilize the biologically active form of the compound. Salts dissolved in buffered solutions are utilized as diluents in the art. One commonly used buffered solution is phosphate buffered saline because it mimics the salt conditions of human blood. Since buffer salts can control the pH of a solution at low concentrations, a buffered diluent rarely modifies the biological activity of a compound.
The term xe2x80x9cpharmaceutical compositionxe2x80x9d refers to a mixture of a compound of the invention with other chemical components, such as diluents, excipients, or carriers. The pharmaceutical composition facilitates administration of the compound to an organism. Multiple techniques of administering a compound exist in the art including, but not limited to, oral, aerosol, parenteral, and topical administration. Pharmaceutical compositions can also be obtained by reacting compounds with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.
The term xe2x80x9cphysiologically acceptablexe2x80x9d defines a carrier or diluent that does not abrogate the biological activity and properties of the compound.
The term xe2x80x9cfunctionxe2x80x9d refers to the cellular role of a protein kinase. The protein kinase family includes members that regulate many steps in signaling cascades, including cascades controlling cell growth, migration, differentiation, gene expression, muscle contraction, glucose metabolism, cellular protein synthesis, and regulation of the cell cycle.
The term xe2x80x9ccatalytic activityxe2x80x9d, in the context of the invention, defines the rate at which a protein kinase phosphorylates a substrate. Catalytic activity can be measured, for example, by determining the amount of a substrate converted to a product as a function of time. Phosphorylation of a substrate occurs at the active-site of a protein kinase. The active-site is normally a cavity in which the substrate binds to the protein kinase and is phosphorylated.
The term xe2x80x9csubstratexe2x80x9d as used herein refers to a molecule phosphorylated by a protein kinase. The substrate is preferably a peptide and more preferably a protein.
The term xe2x80x9cactivatesxe2x80x9d refers to increasing the cellular function of a protein kinase. The protein kinase function is preferably the interaction with a natural binding partner and most preferably catalytic activity.
The term xe2x80x9cinhibitxe2x80x9d refers to decreasing the cellular function of a protein kinase. The protein kinase function is preferably the interaction with a natural binding partner and most preferably catalytic activity.
The term xe2x80x9cmodulatesxe2x80x9d refers to altering the function of a protein kinase by increasing or decreasing the probability that a complex forms between a protein kinase and a natural binding partner. A modulator preferably increases the probability that such a complex forms between the protein kinase and the natural binding partner, more preferably increases or decreases the probability that a complex forms between the protein kinase and the natural binding partner depending on the concentration of the compound exposed to the protein kinase, and most preferably decreases the probability that a complex forms between the protein kinase and the natural binding partner. A modulator preferably activates the catalytic activity of a protein kinase, more preferably activates or inhibits the catalytic activity of a protein kinase depending on the concentration of the compound exposed to the protein kinase, or most preferably inhibits the catalytic activity of a protein kinase.
The term xe2x80x9ccomplexxe2x80x9d refers to an assembly of at least two molecules bound to one another. Signal transduction complexes often contain at least two protein molecules bound to one another.
The term xe2x80x9cnatural binding partnerxe2x80x9d refers to polypeptides that bind to a protein kinase in cells. Natural binding partners can play a role in propagating a signal in a protein kinase signal transduction process. A change in the interaction between a protein kinase and a natural binding partner can manifest itself as an increased or decreased probability that the interaction forms, or an increased or decreased concentration of the protein kinase/natural binding partner complex.
The term xe2x80x9ccontactingxe2x80x9d as used herein refers to mixing a solution comprising a compound of the invention with a liquid medium bathing the cells of the methods. The solution comprising the compound may also comprise another component, such as dimethylsulfoxide (DMSO), which facilitates the uptake of the compound or compounds into the cells of the methods. The solution comprising the compound of the invention may be added to the medium bathing the cells by utilizing a delivery apparatus, such as a pipet-based device or syringe-based device.
The term xe2x80x9cmonitoringxe2x80x9d refers to observing the effect of adding the compound to the cells of the method. The effect can be manifested in a change in cell phenotype, cell proliferation, protein kinase catalytic activity, or in the interaction between a protein kinase and a natural binding partner.
The term xe2x80x9ceffectxe2x80x9d describes a change or an absence of a change in cell phenotype or cell proliferation. xe2x80x9cEffectxe2x80x9d can also describe a change or an absence of a change in the catalytic activity of the protein kinase. xe2x80x9cEffectxe2x80x9d can also describe a change or an absence of a change in an interaction between the protein kinase and a natural binding partner.
The term xe2x80x9ccell phenotypexe2x80x9d refers to the outward appearance of a cell or tissue or the function of the cell or tissue. Examples of cell phenotype are cell size (reduction or enlargement), cell proliferation (increased or decreased numbers of cells), cell differentiation (a change or absence of a change in cell shape), cell survival, apoptosis (cell death), or the utilization of a metabolic nutrient (e.g., glucose uptake). Changes or the absence of changes in cell phenotype are readily measured by techniques known in the art.
The term xe2x80x9cantibodyxe2x80x9d refers to an antibody (e.g., a monoclonal or polyclonal antibody), or antibody fragment, having specific binding affinity to protein kinase or its fragment.
By xe2x80x9cspecific binding affinityxe2x80x9d is meant that the antibody binds to target (protein kinase) polypeptides with greater affinity than it binds to other polypeptides under specified conditions. Antibodies having specific binding affinity to a protein kinase may be used in methods for detecting the presence and/or amount of a protein kinase in a sample by contacting the sample with the antibody under conditions such that an immunocomplex forms and detecting the presence and/or amount of the antibody conjugated to the protein kinase. Diagnostic kits for performing such methods may be constructed to include a first container containing the antibody and a second container having a conjugate of a binding partner of the antibody and a label, such as, for example, a radioisotope. The diagnostic kit may also include notification of an FDA approved use and instructions therefor.
The term xe2x80x9cpolyclonalxe2x80x9d refers to antibodies that are heterogenous populations of antibody molecules derived from the sera of animals immunized with an antigen or an antigenic functional derivative thereof. For the production of polyclonal antibodies, various host animals may be immunized by injection with the antigen. Various adjuvants may be used to increase the immunological response, depending on the host species.
xe2x80x9cMonoclonal antibodiesxe2x80x9d are substantially homogenous populations of antibodies to a particular antigen. They may be obtained by any technique which provides for the production of antibody molecules by continuous cell lines in culture. Monoclonal antibodies may be obtained by methods known to those skilled in the art. See, for example, Kohler, et al., Nature 256:495-497 (1975), and U.S. Pat. No. 4,376,110.
The term xe2x80x9cantibody fragmentxe2x80x9d refers to a portion of an antibody, often the hypervariable region and portions of the surrounding heavy and light chains, that displays specific binding affinity for a particular molecule. A hypervariable region is a portion of an antibody that physically binds to the polypeptide target.
The term xe2x80x9caberrationxe2x80x9d, in conjunction with a signal transduction process, refers to a protein kinase that is over- or under-expressed in an organism, mutated such that its catalytic activity is lower or higher than wild-type protein kinase activity, mutated such that it can no longer interact with a natural binding partner, is no longer modified by another protein kinase or protein phosphatase, or no longer interacts with a natural binding partner.
The term xe2x80x9cpromoting or disrupting the abnormal interactionxe2x80x9d refers to a method that can be accomplished by administering a compound of the invention to cells or tissues in an organism. A compound can promote an interaction between a protein kinase and natural binding partners by forming favorable interactions with multiple atoms at the complex interface. Alternatively, a compound can inhibit an interaction between a protein kinase and natural binding partners by compromising favorable interactions formed between atoms at the complex interface.
xe2x80x9cIn vitroxe2x80x9d refers to procedures performed in an artificial environment, such as, without limitation, in a test tube, in a cell, or culture medium. As used herein, xe2x80x9cin vivoxe2x80x9d refers to procedures performed within a living organism such as, without limitation, a mouse, rat, or rabbit.
In one aspect, the present invention relates to 3-arylidenyl-6-heterocyclyl-2-indolinone derivatives having the chemical structure set forth in formula I: 
or a salt or prodrug thereof where n and m are independently 0 or 1.
When n is 1, then A, B, D, E and F are independently selected from the group consisting of carbon and nitrogen; however, no more than three of A, B, D, E and F are nitrogen at the same time and, when A, B, D, E, or F is nitrogen, then R4, R5, R6, R7 or R8, respectively, does not exist.
When m is 1, then G, H, J, K and L are independently selected from the group consisting of carbon and nitrogen; however, at least one and no more than three of G, H, J, K and L are nitrogen at the same time and, when G, H, J, K or L is nitrogen, then R9, R10, R11, R12 or R13, respectively, does not exist.
When n is 0, then A is selected from the group consisting of carbon and nitrogen, B and F are selected from the group consisting of carbon, nitrogen, NH, oxygen and sulfur, provided that when B or F is NH, the other cannot be NH, and E is selected from the group consisting of carbon, nitrogen, oxygen and sulfur, further provided that no more than one of B, E or F is oxygen or sulfur and provided also that at least one of A, B, E or F is a heteroatom (i.e., not carbon).
When m is 0, then G is selected from the group consisting of carbon and nitrogen, H, K and L are selected from the group consisting of carbon, nitrogen, NH, oxygen and sulfur, provided that when H or L is NH, the other cannot be NH, and K is selected from the group consisting of carbon, nitrogen, oxygen and sulfur, further provided that no more than one of H, K or L is oxygen or sulfur and provided also that at least one of G, H, K or L is a heteroatom (i.e., not carbon).
R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12 and R13 are independently selected from the group consisting of hydrogen, alkyl, trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, mercapto, alkylthio, aryloxy, arylthio, sulfinyl, sulfonyl, S-sulfonamido, N-sulfonamido, carbonyl, C-carboxy, O-carboxy, carboxyalkyl, cyano, nitro, halo, O-carbamyl, N-carbamyl, C-amido, N-amido and xe2x80x94NR14R15.
R4 and R5 or R5 and R6 or R6 and R7 or R8 and R8 may combine to form a five-member or a six-member aryl or heteroaryl ring.
R14 and R15 are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, carbonyl, sulfonyl, and, combined, a five-member or a six-member heteroalicyclic ring.
A presently preferred embodiment of this invention is a compound of formula I wherein,
R1, R2 and R3 are independently selected from the group consisting of (i) hydrogen, (ii) lower alkyl optionally substituted with one or more halo groups, (iii) lower alkoxy optionally substituted with one or more halo groups, (iv) (lower alkyl)-S-sulfonamido, (v) aryl-S-sulfonamido, (vi) halo, and (vii) xe2x80x94NR14R15; and,
R4, R5, R6, R7, R8, R9, R10, R11, R12, and R13 are independently selected from the group consisting of (i) hydrogen; (ii) lower alkyl, optionally substituted with one or more groups selected from the group consisting of aryl, heteroaryl, heteroalicyclic, halo, hydroxy, lower alkoxy, mercapto, lower alkylthio, C-carboxy and xe2x80x94NR14R15; (iii) cycloalkyl; (iv) hydroxy; (v) lower alkoxy, optionally substituted with one or more groups selected from the group consisting of one or more halo groups, aryl, heteroaryl and heteroalicyclic; (vi) halo; (vii) carboxyalkyl; (viii) aryl, optionally substituted with one or more groups selected from the group consisting of lower alkyl optionally substituted one or more halo groups, halo, hydroxy, lower alkoxy optionally substituted with one or more halo groups, aryloxy and xe2x80x94NR14R15; (ix) aryloxy, optionally substituted with one or more groups selected from the group consisitng of lower alkyl optionally substituted with one or more halo groups, halo, hydroxy, lower alkoxy optionally substituted with one or more halo groups, aryloxy and xe2x80x94NR14R15; (x) heteroaryl, optionally substituted with one or more groups selected from the group consisting of lower alkyl optionally substituted with one or more halo groups, halo, hydroxy,. lower alkoxy optionally substituted with one or more halo groups and xe2x80x94NR14R15; (xi) heteroalicyclic; (xii) (lower alkyl)-C-carboxy; (xiii) (lower alkyl)carbonyl; (xiv) aryl carbonyl; (xv) (lower alkyl)-S-sulfonamido; (xvi) aryl-S-sulfonamido; (xvii) (lower alkyl)-N-sulfonamido; (xviii) aryl-N-sulfonamido; (xix) (lower alkyl)-N-carbamoyl; (xx) (lower alkyl)-C-amido; (xxi) (lower alkyl)-N-amido; (xxii) (cycloalkyl)-N-amido; and, (xxiii) xe2x80x94NR14R15; and
R14 and R15 are independently selected from the group consisting of hydrogen and lower alkyl.
Another presently preferred embodiment of this invention is a compound of formula I in which n is 1; A, B, D, E and F are carbon, m is 1 and one of G, H, J, K and L is nitrogen.
It is also a presently preferred embodiment of this invention that, in a compound of formula I, n is 1; A, B, D, E and F are carbon, m is 0, G, H and K are carbon, and L is NH, oxygen or sulfur.
A still further preferred embodiment of this invention is a compound of formula I in which n is 0; A, B and E are carbon; F is NH; R4 and R7 are independently selected from the group consisting of hydrogen and lower alkyl; R5 is selected from the group consisting of (i) lower alkyl optionally substituted with one or more groups selected from the group consisting of, hydroxy, heteroaromatic containing an NH group in the ring, heteroalicyclic containing at least one NH group in the ring, C-carboxy, and, xe2x80x94NR14R15; (ii) carboxyalkyl; (iii) C-carboxy; and, (iv) xe2x80x94NR14R15, wherein R14 and R15 are independently selected from the group consisting of hydrogen, lower alkyl and carbonyl; m is 1; and, one of G, H, I, J, K or L is nitrogen.
It is likewise a presently preferred embodiment of this invention that, in a compound of formula I, n is 0; A, B and E are carbon; F is NH; R4 and R7 are independently selected from the group consisting of hydrogen and lower alkyl; R5 is selected from the group consisting of (i) lower alkyl optionally substituted with one or more groups selected from the group consisting of, hydroxy, heteroaromatic containing an NH in the ring, heteroalicyclic containing at least one NH group in the ring, C-carboxy, and, xe2x80x94NR14R15; (ii) carboxyalkyl; (iii) C-carboxy; and, (iv) xe2x80x94NR14R15, wherein R14 and R15 are independently selected from the group consisting of hydrogen, lower alkyl and carbonyl; m is 0; G, H and K are carbon; and, L is NH, oxygen or sulfur.
In another aspect, the present invention relates to 3-aralkyl-2-indolinone derivatives having the chemical structure set forth in formula II: 
or a physiologically acceptable salt or prodrug thereof where P is 0 or 1.
When p is 1, then M, Q, T, U and V are independently selected from the group consisting of carbon and nitrogen, it being understood that, when M, Q, T, U, or V is nitrogen, R20, R21, R22, R23, or R24, respectively, do not exist.
When p is 0, then M, Q, U, and V are independently selected from the group consisting of carbon, nitrogen, oxygen and sulfur, it being understood that, when M, Q, U, or V is oxygen or sulfur or nitrogen (wherein said nitrogen is participating in a double bond), R20, R21, R22, R23, or R24, respectively, do not exist.
R16, R17, R18, R19, R20, R21, R22, R23, or R24 are independently selected from the group consisting of hydrogen, alkyl, trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, mercapto, alkylthio, aryloxy, sulfinyl, sulfonyl, S-sulfonamido, N-sulfonamido, carbonyl, C-carboxy, O-carboxy, carboxyalkyl, cyano, nitro, halo, O-carbamyl, N-carbamyl, C-amido, N-amido and xe2x80x94NR25R26.
R20 and R21 or R21 and R22 or R23 and R23 or R23 and R24 may combine to form a five-member or a six-member aryl or heteroaryl ring.
R25 and R26 are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, carbonyl, sulfonyl, and, combined, a five-member or a six-member heteroalicyclic ring.
A presently preferred embodiment of this invention is a compound wherein R16, R17, R18, R19, R20, R21, R22, R23, and R24 are independently selected from the group consisting of (i) hydrogen; (ii) lower alkyl, optionally substituted with one or more groups selected from the group consisting of cycloalkyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, C-carboxy and xe2x80x94NR25R26; (iii) cycloalkyl; (iv) hydroxy; (v) lower alkoxy,optionally substituted with one or more groups selected from the group consisting of one or more halo groups, aryl, heteroaryl and heteroalicyclic; (vi) trihalomethyl; (vii) trihalomethoxy; (viii) halo; (ix) carboxyalkyl; (x) aryl,optionally substituted with one or more groups selected from the group consisting of lower alkyl, lower alkyl substituted with one or more halo groups, trihalomethyl, trihalomethoxy, halo, hydroxy, lower alkoxy, aryloxy and xe2x80x94NR25R26; (xi) aryloxy; heteroaryl, optionally substituted with one or more groups selected from the group consisting of lower alkyl, trihalomethyl, halo, hydroxy, (lower alkyl)alkoxy, amino and xe2x80x94NR25R26; (xii) heteroalicyclic; (xiii) (lower alkyl)carboxy; (xiv) (lower alkyl)carbonyl; (xv) aryl carbonyl; (xvi) (lower alkyl)-S-sulfonamido; (xvii) aryl-S-sulfonamido; (xviii) (lower alkyl)-N-sulfonamido; (xix) aryl-N-sulfonamido; (xx) (lower alkyl)-N-carbamoyl; (xxi) (lower alkyl)-C-amido; (xxii) (lower alkyl)-N-amido; (xxiii) (cycloalkyl)-N-amido; and, (xxiv) xe2x80x94NR25R26.
Another presently preferred embodiment of this invention is a compound wherein p is 1 and M, Q, T, U, and V are carbon.
It is a further presently preferred embodiment of this invention that, when p is 1 and M, Q, T, U, and V are carbon; R16, R17, R18, and R19 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) S-sulfonamido optionally substituted with one or more groups selected from the group consisting of hydrogen, lower alkyl and aryl; (vi) lower alkyl optionally substituted with a group selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic, and xe2x80x94NR25R26; (vii) lower alkoxy optionally substituted with one or more halo groups; (viii) aryl optionally substituted with one or more groups selected from the group consisting of lower alkyl, lower alkoxy, halo, hydroxy and xe2x80x94NR25R26; (ix) N-amido optionally substituted with one or more groups selected from the group consisting of hydrogen, lower alkyl, cycloalkyl and aryl; and R20, R21, R22, R23, and R24 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) lower alkyl optionally substituted with one or more groups selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic, and xe2x80x94NR25R26; (vi) cycloalkyl; (vii) lower alkoxy optionally substituted with one or more groups selected from the group consisting of one or more halo groups, aryl, xe2x80x94NR25R26, and heteroaryl, (viii) aryl optionally substituted with one or more groups selected from the group consisting of halo, hydroxy, lower alkoxy, xe2x80x94NR25R26, and C-carboxy.
It is yet another presently preferred embodiment of this invention that p is 1 and one or two of M, Q, T, U, or V are nitrogen.
It is likewise presently preferred that, when p is 1 and one or two of, Q, T, U, and V are nitrogen, R16, R17, R18, and R19 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) S-sulfonamido optionally substituted with one or more groups selected from the group consisting of hydrogen, lower alkyl and aryl; (vi) lower alkyl optionally substituted with a group selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic, and xe2x80x94NR25R26; (vii) lower alkoxy optionally substituted with one or more halo groups; (viii) aryl optionally substituted with one or more groups selected from the group consisting of lower alkyl, lower alkoxy, halo, hydroxy and xe2x80x94NR25R26; (ix) N-amido optionally substituted with one or more groups selected from the group consisting of hydrogen, lower alkyl, cycloalkyl and aryl; and, R20, R21, R22, R23, and R24, whichever of these are not nitrogen, are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) lower alkyl optionally substituted with one or more groups selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic, and xe2x80x94NR25R26; (vi) cycloalkyl; (vii) lower alkoxy optionally substituted with one or more groups selected from the group consisting of one or more halo groups, aryl, xe2x80x94NR25R26, and heteroaryl; and (viii) aryl optionally substituted with one or more groups selected from the group consisting of halo, hydroxy, lower alkoxy, xe2x80x94NR25R26, and C-carboxy.
It is a presently preferred embodiment of this invention that p is 1 and R21, and R22 combine to form a fused pyrrolo group.
In further preferred embodiments of the invention, R16, R17, R18, and R19 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) S-sulfonamido optionally substituted with one or more groups selected from the group consisting of hydrogen, lower alkyl and aryl; (vi) lower alkyl optionally substituted with a group selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic and, xe2x80x94NR25R26; (vii) lower alkoxy optionally substituted with one or more halo groups; (viii) aryl optionally substituted with one or more groups selected from the group consisting of lower alkyl, lower alkoxy, halo, hydroxy and xe2x80x94NR25R26; (ix) N-amido optionally substituted with one or more groups selected from the group consisting of hydrogen, lower alkyl, cycloalkyl and aryl.
Similarly it is preferred that R20, R23, and R24 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) lower alkyl optionally substituted with one or more groups selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic and, xe2x80x94NR25R26; (vi) cycloalkyl; (vii) lower alkoxy optionally substituted with one or more groups selected from the group consisting of one or more halo groups, aryl, xe2x80x94NR25R26 and, heteroaryl; (viii) aryl optionally substituted with one or more groups selected from the group consisting of halo, hydroxy, lower alkoxy, xe2x80x94NR25R26 and, C-carboxy.
In another preferred embodiment, p is 0, M or Q is xe2x80x94NH, oxygen or sulfur; and, M or Q, whichever is not xe2x80x94NH, oxygen or sulfur, and U and V are carbon.
Other preferred embodiments include compounds in which R16, R17, R18, and R19 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) S-sulfonamido optionally substituted with one or more groups selected from the group consisting of hydrogen, lower alkyl and aryl; (vi) lower alkyl optionally substituted with a group selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic and, xe2x80x94NR25R26; (vii) lower alkoxy optionally substituted with one or more halo groups; (viii) aryl optionally substituted with one or more groups selected from the group consisting of lower alkyl, lower alkoxy, halo, hydroxy and xe2x80x94NR25R26; (ix) N-amido optionally substituted with one or more groups selected from hydrogen, lower alkyl, cycloalkyl and aryl.
In these preferred compounds R20, R21, R23, and R24 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) lower alkyl optionally substituted with one or more groups selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic and, xe2x80x94NR25R26; (vi) cycloalkyl; (vii) lower alkoxy optionally substituted with one or more groups selected from the group consisting of one or more halo groups, aryl, xe2x80x94NR25R26, and, heteroaryl.
In still further preferred embodiments, R16, R17, R18, and R19 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) S-sulfonamido optionally substituted with one or more groups selected from the group consisting of hydrogen, lower alkyl and aryl; (vi) lower alkyl optionally substituted with a group selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic and, xe2x80x94NR25R26; (vii) lower alkoxy optionally substituted with one or more halo groups; (viii) aryl optionally substituted with one or more groups selected from the group consisting of lower alkyl, lower alkoxy, halo, hydroxy and xe2x80x94NR25R26; (ix) N-amido optionally substituted with one or more groups selected from hydrogen, lower alkyl, cycloalkyl and aryl.
Preferably, R20, R23, and R24 are independently selected from the group consisting of (i) hydrogen; (ii) halo; (iii) hydroxy; (iv) xe2x80x94NR25R26; (v) lower alkyl optionally substituted with one or more groups selected from the group consisting of hydroxy, one or more halo groups, C-carboxy, C-amido, heteroalicyclic and, xe2x80x94NR25R26; (vi) cycloalkyl; (vii) lower alkoxy optionally substituted with one or more groups selected from the group consisting of one or more halo groups, aryl, xe2x80x94NR25R26 and, heteroaryl; (viii) aryl optionally substituted with one or more groups selected from the group consisting of halo, hydroxy, lower alkoxy, xe2x80x94NR25R26 and, C-carboxy.
In another aspect, the invention provides a compound having a structure set forth in formula III 
where
(a) is selected from the group consisting of
(i) saturated or unsaturated alkyl optionally substituted with substituents selected from the group consisting of halogen, trihalomethyl, alkoxy, carboxylate, amino, nitro, ester, and a five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moiety,
xe2x80x83where the ring moiety is optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties; and
(ii) an aromatic or heteroaromatic ring optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iii) an aliphatic or heteroaliphatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, ester, and an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(b) R32, R33, and R34 are each independently selected from the group consisting of
(i) hydrogen;
(ii) saturated or unsaturated alkyl optionally substituted with one or more substituents selected from the group consisting of halogen, trihalomethyl, carboxylate, amino, nitro, ester, and a five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moiety,
xe2x80x83where the ring moiety is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iii) an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iv) an aliphatic or heteroaliphatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, ester, and an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(v) an amine of formula xe2x80x94(X1)n1xe2x80x94NX2X3, where
xe2x80x83X1 is selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, or aliphatic ring moieties,
xe2x80x83n1 is 0 or 1, and
xe2x80x83X2 and X3 are independently selected from the group consisting of hydrogen, saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, or aliphatic ring moieties;
(vi) a nitro of formula xe2x80x94NO2;
(vii) a halogen or trihalomethyl;
(viii) a ketone of formula xe2x80x94(X4)n4xe2x80x94COxe2x80x94X5, where
xe2x80x83X4 and X5 are independently selected from the group consisting of alkyl and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties and where, and
xe2x80x83n4 is 0 or 1;
(ix) a carboxylic acid of formula xe2x80x94(X6)n6xe2x80x94COOH or ester of formula xe2x80x94(X7)n7xe2x80x94COOxe2x80x94X8, where
xe2x80x83X6, X7, and X8 and are independently selected from the group consisting of alkyl and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties, and
xe2x80x83n6 and n7 are independently 0 or 1;
(x) an alcohol of formula xe2x80x94(X9)n9xe2x80x94OH or an alkoxyalkyl moiety of formula xe2x80x94(X10)n10xe2x80x94Oxe2x80x94X11, where
xe2x80x83X9, X10, and X11 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n9 and n10 are independently 0 or 1;
(xi) an amide of formula xe2x80x94(X12)n12xe2x80x94NHCOX13, or of formula xe2x80x94(X14)n14xe2x80x94CONX15X16, where
xe2x80x83X12 and X14 are each independently selected from the group consisting of alkyl, hydroxyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the ring is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n12 and n14 are independently 0 or 1,
xe2x80x83X13, X15, and X16 are each independently selected from the group consisting of hydrogen, alkyl, hydroxyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the ring is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester;
(xii) a sulfonamide of formula xe2x80x94(X17)n17xe2x80x94SO2NX18X19, where
xe2x80x83X17 is selected from the group consisting of alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester,
xe2x80x83n17 is 0, 1, or 2,
xe2x80x83X18, and X19 are independently selected from the group consisting of hydrogen, alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, or
xe2x80x83where X18 and X19 taken together form a five-membered or six-membered aliphatic or heteroaliphatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester;
(xiii) an aldehyde of formula xe2x80x94(X20)n20xe2x80x94COxe2x80x94H where
xe2x80x83X20 is selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n20 is 0 or 1;
(xiv) a sulfone of formula xe2x80x94(X21)n21xe2x80x94SO2xe2x80x94X22, where
xe2x80x83X21 and X22 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n21 is 0 or 1; and
(xv) a thiol of formula xe2x80x94(X23)n23xe2x80x94SH or a thioether of formula xe2x80x94(X24)n24xe2x80x94Sxe2x80x94X25, where
xe2x80x83X23, X24, and X25 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n23 and n24 are independently 0 or 1; or
xe2x80x83R33 and R34 taken together form a six-membered aliphatic or aromatic ring, optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester; and
(c) R28, R29, R30, and R31 are each independently selected from the group consisting of
(i) hydrogen;
(ii) saturated or unsaturated alkyl optionally substituted with one or more substituents selected from the group consisting of hydroxy, halogen, trihalomethyl, carboxylate, amino, nitro, ester, and a five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moiety,
xe2x80x83the ring moiety is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iii) an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iv) an aliphatic or heteroaliphatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, ester, and an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(v) an amine of formula xe2x80x94(X1)n1xe2x80x94NX2X3, where
xe2x80x83X1 is selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, or aliphatic ring moieties,
xe2x80x83n1 is 0 or 1, and
xe2x80x83X2, and X3 are independently selected from the group consisting of hydrogen, saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, or aliphatic ring moieties;
(vi) a nitro of formula xe2x80x94NO2;
(vii) a halogen or trihalomethyl;
(viii) a ketone of formula xe2x80x94(X4)n4xe2x80x94COxe2x80x94X5, where
xe2x80x83X4 and X5 are independently selected from the group consisting of alkyl and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties, and
xe2x80x83n4 is 0 or 1;
(ix) a carboxylic acid of formula xe2x80x94(X6)n6xe2x80x94COOH or ester of formula xe2x80x94(X7)n7xe2x80x94COOxe2x80x94X8, where
xe2x80x83X6, X7, and X8 and are independently selected from the group consisting of alkyl and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties, and n6 and n7 are independently 0 or 1;
(x) an alcohol of formula xe2x80x94(X9)n9xe2x80x94OH or an alkoxyalkyl moiety of formula xe2x80x94(X10)n10xe2x80x94Oxe2x80x94X11 where
xe2x80x83X9, X10, and X11 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n9 and n10 are independently 0 or 1;
(xi) an amide of formula xe2x80x94(X12)n2xe2x80x94NHCOX13, or of formula xe2x80x94(X14)n14xe2x80x94CONX15X16, where
xe2x80x83X12 and X14 are each independently selected from the group consisting of alkyl, hydroxyl, and five-membered or six-membered aromatic,
xe2x80x83heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester,
xe2x80x83n12 and n14 are independently 0 or 1, and
xe2x80x83X13, X15, and X16 are each independently selected from the group consisting of hydrogen, alkyl, hydroxyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties, and
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester;
(xii) a sulfonamide of formula xe2x80x94(X17)n17xe2x80x94SO2NX18X19, where
xe2x80x83X17 is selected from the group consisting of alkyl, five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n17 is 0, 1, or2, and
xe2x80x83X18, and X19 are independently selected from the group consisting of hydrogen, alkyl, five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, or
xe2x80x83X18 and X19 taken together form a five-membered or six-membered aliphatic or heteroaliphatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester;
(xiii) an aldehyde of formula xe2x80x94(X20)n20xe2x80x94COxe2x80x94H where
xe2x80x83X20 is selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and n20 is 0or 1;
(xiv) a sulfone of formula xe2x80x94(X21)n21xe2x80x94SO2xe2x80x94X22, where
xe2x80x83X21 and X22 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n21 is 0 or 1; and
(xv) a thiol of formula xe2x80x94(X23)n23xe2x80x94SH or a thioether of formula xe2x80x94(X24)n24xe2x80x94Sxe2x80x94X25, where
xe2x80x83X23, X24, and X25 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n23 and n24 are independently 0 or 1.
In preferred embodiments, the invention relates to a compound of formula III where
(a) R31 is hydrogen;
(b) R32, R33, and R34 are each independently selected from the group consisting of
(i) hydrogen;
(ii) saturated alkyl optionally substituted with a six-membered heteroaliphatic ring moiety;
(iii) an amine of formula xe2x80x94(X1)n1xe2x80x94NX2X3, where X1 is saturated alkyl, n1 is 0 or 1, and X2 and X3 are independently selected from the group consisting of hydrogen and saturated alkyl;
(iv) a carboxylic acid of formula xe2x80x94(X6)n6xe2x80x94COOH or ester of formula xe2x80x94(X7)n7xe2x80x94COOxe2x80x94X8, where X6, X7, and X8 are alkyl, and n6 and n7 are independently 0 or 1; and
(v) an amide of formula xe2x80x94(X14)n14xe2x80x94CONX15X16, where X4 is alkyl, n12 and n14 are independently 0 or 1, and X15 and X16 are each independently selected from the group consisting of hydrogen and alkyl;
xe2x80x83or R7 is as described herein and R8 and R9 taken together form an optionally substituted six-membered aliphatic or aromatic ring; and
(c) R28, R29, and R30 are each independently selected from the group consisting of
(i) hydrogen;
(ii) saturated alkyl;
(iii) a halogen or trihalomethyl;
(iv) a carboxylic acid of formula xe2x80x94(X6)n6xe2x80x94COOH or ester of formula xe2x80x94(X7)n7xe2x80x94COOxe2x80x94X8, where X6, X7, and X8 and are alkyl, and n6 and n7 are independently 0 or 1;
(v) an alcohol of formula xe2x80x94(X9)n9xe2x80x94OH or an alkoxyalkyl moiety of formula xe2x80x94(X10)n10xe2x80x94Oxe2x80x94X11, where X9, X10, and X11 are alkyl, and n9 and n10 are independently 0 or 1;
(vi) an amide of formula xe2x80x94(X12)n12xe2x80x94NHCOX13, or of formula xe2x80x94(X14)n14xe2x80x94CONX15X16, where X12 and X14 are alkyl, n12 and n14 are independently 0 or 1, and X13, X15, and X16 are each independently selected from the group consisting of hydrogen and alkyl; and
(vii) a sulfonamide of formula xe2x80x94(X17)n17xe2x80x94SO2NX18X19, where X17 is alkyl, and n17 is 0, 1, or 2, and X18, and X19 are independently selected from the group consisting of hydrogen and alkyl.
More preferably, in the compounds of formula III
(a) R28 is selected from the group consisting of hydrogen, methyl, and 2-hydroxyethyl;
(b) R29 is selected from the group consisting of hydrogen, methyl, chloro, bromo, carboxy, methoxy, xe2x80x94NHC(O)xe2x80x94CH3, and xe2x80x94SO2N(CH3)2;
(c) R30 is selected from the group consisting of hydrogen, chloro, methoxy, and xe2x80x94NHC(O)xe2x80x94CH3;
(d) R31 is hydrogen;
(e) R32 is selected from the group consisting of hydrogen, methyl, xe2x80x94CH2CH2C(O)OH, xe2x80x94CH2CH2C(O)NH2, xe2x80x94CH2CH2CH2N(CH3)2, and 3-morpholinopropyl; and
(f) R33 and R34 are each independently selected from the group consisting of hydrogen, methyl, xe2x80x94CH2CH2C(O)OH, xe2x80x94CH2CH2CH2N(CH3)2, and 3-morpholinopropyl, or R8 and R9 taken together form a six-membered aromatic or aliphatic ring.
The preferred diaryl indolinone compounds of the invention are those which are preferrably formed by the reaction of a ketone compound with and oxindole compound. The ketone compound is preferrably selected from the group consisting of 
In the above structures R27 is selected from the group consisting of
(i) saturated or unsaturated alkyl optionally substituted with substituents selected from the group consisting of halogen, trihalomethyl, alkoxy, carboxylate, amino, nitro, ester, and a five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moiety,
xe2x80x83wherein said ring moiety is optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties; and
(ii) an aromatic or heteroaromatic ring optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iii) an aliphatic or heteroaliphatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, ester, and an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties.
The oxindole compound is preferrably selected from the group consisting of 1,3-dihydro-indol-2-one, 4-methyl-1,3-dihydro-indol-2-one, 4-(2-hydroxy-ethyl)-1,3-dihydro-indol-2-one, 5-chloro-1,3-dihydro-indol-2-one, 5-bromo-1,3-dihydro-indol-2-one, 5-methoxy-1,3-dihydro-indol-2-one, 2-oxo-2,3-dihydro-1H-indole-5-carboxylic acid, 2-oxo-2,3-dihydro-1H-indole-5-sulfonic acid dimethylamide, N-(2-oxo-2,3-dihydro-1H-indole-5-yl)-acetamide, 6-chloro-1,3-dihydro-indol-2-one, 6-methoxy-1,3-dihydro-indol-2-one, and N-(5-methyl-2-oxo-2,3-dihydro-1H-indole-6-yl)-acetamide.
In a further aspect, the invention provides a compound having a structure set forth in formula IV or formula V: 
where
(a) R35, R36, and R41 are each independently selected from the group consisting of
(i) hydrogen;
(ii) saturated or unsaturated alkyl optionally substituted with substituents selected from the group consisting of halogen, trihalomethyl, alkoxy, carboxylate, amino, nitro, ester, and a five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moiety,
xe2x80x83where the ring moiety is optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties; and
(iii) an aromatic or heteroaromatic ring optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(b) R37 is an ethyl-2-oxy group of formula xe2x80x94CH2CH2xe2x80x94Oxe2x80x94R, where R is selected from the group consisting of
(i) hydrogen;
(ii) saturated or unsaturated alkyl optionally substituted with substituents selected from the group consisting of halogen, trihalomethyl, carboxylate, amino, nitro, ester, and a five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moiety, and
xe2x80x83the ring moiety is optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iii) an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, aryl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties
(iv) a substituent of formula xe2x80x94C(E)NX15X16, where
xe2x80x83X15, and X16 are each independently selected from the group consisting of hydrogen, alkyl, hydroxyl, sulfone of formula xe2x80x94SO2xe2x80x94X22, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the ring is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, aryl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester,
xe2x80x83X22 is selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the ring is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester,
xe2x80x83E is selected from the group consisting of oxygen and sulfur; and
(c) R38, R39, and R40 are each hydrogen;
(d) Z is a 5, 6, 7, 8, 9, or 10 membered, monocyclic or bicyclic, aromatic or heteroaromatic, ring moiety, optionally substituted with one or more substituents selected from the group consisting of
(i) hydrogen;
(ii) saturated or unsaturated alkyl optionally substituted with one or more substituents selected from the group consisting of hydroxy, halogen, trihalomethyl, carboxylate, amino, nitro, ester, and a five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moiety,
xe2x80x83the ring moiety is optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iii) an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(iv) an aliphatic or heteroaliphatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, ester, and an aromatic or heteroaromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkoxy, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties;
(v) an amine of formula xe2x80x94(X1)n1xe2x80x94NX2X3, where
xe2x80x83X1 is selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, or aliphatic ring moieties,
xe2x80x83n1 is 0 or 1, and
xe2x80x83X2, and X3 are independently selected from the group consisting of hydrogen, saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, or aliphatic ring moieties;
(vi) a nitro of formula xe2x80x94NO2;
(vii) a halogen or trihalomethyl;
(viii) a ketone of formula xe2x80x94(X4)n4xe2x80x94COxe2x80x94X5, where X4 and X5 are independently selected from the group consisting of alkyl and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester moieties, and
xe2x80x83n4 is 0 or 1;
(ix) a carboxylic acid of formula xe2x80x94(X6)n6xe2x80x94COOH or ester of formula xe2x80x94(X7)n7xe2x80x94COOxe2x80x94X8, where
xe2x80x83X6, X7, and X8 and are independently selected from the group consisting of alkyl and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties, and n6 and n7 are independently 0 or 1;
(x) an alcohol of formula xe2x80x94(X9)n9xe2x80x94OH or an alkoxyalkyl moiety of formula xe2x80x94(X10)n10xe2x80x94Oxe2x80x94X11, where
xe2x80x83X9, X10, and X11 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n9 and n 10 are independently 0 or 1;
(xi) an amide of formula xe2x80x94(X12)n12xe2x80x94NHCOX13, or of formula xe2x80x94(X14)n14xe2x80x94CONX15X16, where
xe2x80x83X12 and X14 are each independently selected from the group consisting of alkyl, hydroxyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester,
xe2x80x83n12 and n14 are independently 0 or 1, and
xe2x80x83X13, X15, and X16 are each independently selected from the group consisting of hydrogen, alkyl, hydroxyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties, and
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester;
(xii) a sulfonamide of formula xe2x80x94(X17)n17xe2x80x94SO2NX18X19, where
xe2x80x83X17 is selected from the group consisting of alkyl, five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, or ester, and
xe2x80x83n17 is 0, 1, or 2, and
xe2x80x83X18, and X19 are independently selected from the group consisting of hydrogen, alkyl, five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, or ester, or
xe2x80x83X18 and X19 taken together form a five-membered or six-membered aliphatic or heteroaliphatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester;
(xiii) an aldehyde of formula xe2x80x94(X20)n20xe2x80x94COxe2x80x94H where
xe2x80x83X20 is selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n20 is 0 or 1;
(xiv) a sulfone of formula xe2x80x94(X21)n21xe2x80x94SO2xe2x80x94X22, where
xe2x80x83X21 and X22 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n21 is 0 or 1; and
(xv) a thiol of formula xe2x80x94(X23)n23xe2x80x94SH and a thioether of formula xe2x80x94(X24)n24xe2x80x94Sxe2x80x94X25, where
xe2x80x83X23, X24, and X25 are independently selected from the group consisting of saturated or unsaturated alkyl, and five-membered or six-membered aromatic, heteroaromatic, aliphatic, or heteroaliphatic ring moieties,
xe2x80x83the alkyl and ring moieties are optionally substituted with one or more substituents independently selected from the group consisting of alkyl, halogen, trihalomethyl, carboxylate, amino, nitro, and ester, and
xe2x80x83n23 and n24 are independently 0 or 1.
In preferred embodiments, the invention relates to a compound of formula IV or formula V, where
(a) R35, R36, and R41 are hydrogen;
(b) R37 is an ethyl-2-oxy group of formula xe2x80x94CH2CH2xe2x80x94Oxe2x80x94R, where R is selected from the group consisting of hydrogen, saturated alkyl, an aromatic ring optionally substituted with one or more substituents independently selected from the group consisting of alkyl, aryl, and alkoxy moieties, and a substituent of formula xe2x80x94C(E)NHX15, where
xe2x80x83X15 is selected from the group consisting of alkyl, sulfone of formula xe2x80x94SO2xe2x80x94X22, and six-membered aromatic or aliphatic ring moieties,
xe2x80x83the ring is optionally substituted with one or more substituents independently selected from the group consisting of alkyl and aryl, X22 is selected from the group consisting of saturated alkyl, and optionally substituted six-membered aromatic ring moieties, and
xe2x80x83E is selected from the group consisting of oxygen and sulfur, and;
(c) Z is a 5, 6, or 9 membered, monocyclic or bicyclic, aromatic or heteroaromatic, ring moiety, optionally substituted with one or more substituents selected from the group consisting of
(i) hydrogen;
(ii) saturated alkyl,
(iii) an optionally substituted aromatic or heteroaromatic ring;
(iv) an amine of formula xe2x80x94(X1)n1xe2x80x94NX2X3, where
xe2x80x83X1 is alkyl,
xe2x80x83n1 is 0 or 1, and
xe2x80x83X2 and X3 are independently selected from the group consisting of hydrogen and saturated alkyl;
(v) a halogen or trihalomethyl;
(vi) a carboxylic acid of formula xe2x80x94(X6)n6xe2x80x94COOH, where
xe2x80x83X6 is alkyl, and
xe2x80x83n6 is 0 or 1; and
(vii) an alcohol of formula xe2x80x94(X9)n9xe2x80x94OH or an alkoxyalkyl moiety of formula xe2x80x94(X10)n10xe2x80x94Oxe2x80x94X11, where
xe2x80x83X9, X10, and X11 are alkyl, and
xe2x80x83n9 and n10 are independently 0 or 1.
More preferably, in the compounds of formula IV or formula V, R37 is an ethyl-2-oxy group of formula xe2x80x94CH2CH2xe2x80x94Oxe2x80x94R, wherein R is selected from the group consisting of hydrogen, methyl, ethyl, 2-isopropylphenyl, 3-isopropylphenyl, 4-isopropylphenyl, 4-methoxyphenyl, biphen-3-yl, 5-chloropyridin-3-yl, ethylcarbamyl, tert-butylcarbamyl, cyclohexylcarbamyl, phenylcarbamyl, benzene sulfonylcarbamyl, biphen-2-yl-carbamyl, and phenylthiocarbamyl, and Z is selected from the group consisting of 4-bromophenyl, 2-pyridyl, 6-methyl-2-pyridyl, 1H-indol-5-yl, 4-methoxy-3-thiophenphenyl, 4-(3-dimethylamino)-3,5-dimethyl-1H-pyrrol-2-yl, 3-hydroxy-6-methyl-pyridin-2-yl, 4-(3-dimethylaminopropyl)-3,5-dimethyl-1H-pyrrol-2-yl, 3-carboxy-2,4-dimethyl-1H-pyrrol-2-yl, and isopropylcarbamyl.
The preferred compounds of the invention are listed in Table 1.
Some of the above compounds have the structure of formula VI, with the R substituent as defined in Table 2.
Other compounds of the invention have the structure set forth in formula VII, with the substituents as defined in Table 3.
To synthesize the compounds of the invention a base may be used. The base is preferably a nitrogen base or an inorganic base. xe2x80x9cNitrogen basesxe2x80x9d are commonly used in the art and are selected from acyclic and cyclic amines. Examples of nitrogen bases include, but are not limited to, ammonia, methylamine, trimethylamine, triethylamine, aniline, 1,8-diazabicyclo[5.4.0]undec-7-ene, diisopropylethylamine, pyrrolidine, piperidine, and pyridine or substituted pyridine (e.g., 2,6-di-tertbutylpyridine). xe2x80x9cInorganic basesxe2x80x9d are bases that do not contain any carbon atoms. Examples of inorganic bases include, but are not limited to, hydroxide, phosphate, bisulfate, hydrosulfide, and amide anions. Those skilled in the art know which nitrogen base or inorganic base would match the requirements of the reaction conditions. In certain embodiments of the invention, the base used may be pyrrolidine or piperidine. In other embodiments the base may be the hydroxide anion, preferably used as its sodium or potassium salt.
The synthesis of the compounds of the invention may take place in a solvent. The solvent of the reaction is preferably a protic solvent or an aprotic solevent. xe2x80x9cProtic solventsxe2x80x9d are those that are capable of donating a proton to a solute. Examples of protic solvents include, but are not limited to, alcohols and water. xe2x80x9cAprotic solventsxe2x80x9d are those solvents that, under normal reaction conditions, do not donate a proton to a solute. Typical organic solvents, such as hexane, toluene, benzene, methylene chloride, dimethylformamide, chloroform, tetrahydrofuran, are some of the examples of aprotic solvents. Other aprotic solvents are also within the scope used by the present invention. In some preferred embodiments, the solvent of the reaction is an alcohol, which may preferably be isopropanol or most preferably ethanol. Water is another preferred protic solvent. Dimethylformamide, known in the chemistry art as DMF, is a preferred aprotic solvent.
The synthetic method of the invention calls for the reaction to take place at elevated temperatures which are temperatures that are greater than room temperature. More preferably, the elevated temperature is preferably about 30-150xc2x0 C., more preferably about 80-100xc2x0 C., and most preferably about 80-90xc2x0 C., which is about the temperature at which ethanol boils (i.e., the boiling point of ethanol). By xe2x80x9caboutxe2x80x9d a certain temperature it is meant that the temperature range is preferably within 10xc2x0 C. of the listed temperature, more preferably within 5xc2x0 C. of the listed temperature, and most preferably within 2xc2x0 C. of the listed temperature. Therefore, by way of example, by xe2x80x9cabout 80xc2x0 C.xe2x80x9d it is meant that the temperature range is preferably 80xc2x110xc2x0 C., more preferably 80xc2x15xc2x0 C., and most preferably 80xc2x12xc2x0 C.
The synthetic method of the invention may be accompanied by the step of screening a library for a compound of the desired activity and structurexe2x80x94thus, providing a method of synthesis of a compound by first screening for a compound having the desired properties and then chemically synthesizing that compound.
An additional aspect of this invention is a combinatorial library of at least ten 3-arylidenyl-6-heterocyclyl-2-indolinone compounds that can be formed by condensing oxindoles of structure 2 with aldehydes of structure 3.
As used herein, xe2x80x9ccondensingxe2x80x9d or xe2x80x9ccondensationxe2x80x9d refers to a reaction by which two molecules are combined to give one molecule. In particular with regard the present invention a condensation refers to the reaction shown in Scheme I. 
The oxindole in the above combinatorial library is preferrably selected from the group consisting of 6-(pyridin-2-yl)-2-oxindole, 6-(pyridin-3-yl)-2-oxindole, 6-(pyridine-4-yl)-2-oxindole, 6-(pyrimidin-2-yl)-2-oxindole, 6-(pyrimidin-4-yl)-2-oxindole, 6-(pyrimidin-5-yl)-2-oxindole, 6-(triazinyl)-2-oxindole, 6-(pyrrol-2-yl)-2-oxindole, 6-(pyrrol-3-yl)-2-oxindole, 6-(thiophen-2-yl)-2-oxindole, 6-(thiophen-3-yl)-2-oxindole, 6-(furan-2-yl)-2-oxindole, 6-(furan-3-yl)-2-oxindole, 6-(imidazol-2-yl)-2-oxindole, 6-(imidazol-4-yl)-2-oxindole, 6-(thiazol-2-yl)-2-oxindole, 6-(thiazol-4-yl)-2-oxindole, 6-(oxazol-2-yl)-2-oxindole, 6-(oxazol-4-yl)-2-oxindole, 6-(thiadiazol-2-yl)-2-oxindole, 6-(oxadiazolyl)-2-oxindole and 6-(triazol-2-yl)-2-oxindole, 6-(3methylisoxazole-5-yl)-2-oxindole,6-(3-methylisothiazole-5-yl)-2-oxindole, 6-(3,5-dimethyl-isoxazole-4-yl)-2-oxindole, 6-(thiazole-2-yl)-2-oxindole, 6-(thiazole-4-yl)-2-oxindole, 6-(thiazole-5-yl)-2-oxindole, 6-(3-methylthiophene-2-yl)-2-oxindole, 6-(4-methylthiophene-2-yl)-2-oxindole, 6-(5-methylthiophene-2-yl)-2-oxindole, 6-(5-chlorothiophene-2-yl)-2-oxindole, 6-(4-methylfuran-2-yl)-2-oxindole
The aldehyde in the above combinatorial library is preferably selected from the group consisting of, without limitation, benzaldehyde, 3-isopropyl-p-anisaldehyde, 2-methyl-5-isopropyl-p-anisaldehyde, 3,5-diisopropyl-p-anisaldehyde, 3-cyclopentyl-p-anisaldehyde, 3-cyclohexyl-p-anisaldehyde, 3-phenyl-p-anisaldehyde, 3,5-dimethyl-p-anisaldehyde, 2-hydroxy-3,5-dichlorobenzaldehyde, 2-hydroxy-5-chlorobenzaldehyde, 2-hydroxy-4-methoxy-5-(4-methoxyphenyl)benzaldehyde, 3-cyclopentyl-4-methoxybenzaldehyde, 3-cyclopentyl-4-hydroxybenzaldehyde, 3-(2-thienyl)-4-methoxybenzaldehyde, 2-hydroxybenzaldehyde, 2-hydroxy-4-methoxybenzaldehyde, 2-hydroxy-5-bromobenzaldehyde, 2-methylpyridine-6-carboxaldehyde, 3-tert-butyl-4-hydroxy-5-bromobenzaldehyde, 3-bromobenzaldehyde, 3,5-diisopropyl-4-[2-(N-morpholino)ethyl]benzaldehyde,indole-5-carboxaldchyde, 2-hydroxy-3-fluorobenzaldehyde, 3-cyclohexyl-4-[2-(N-morpholino)ethyl]benzaldehyde, 3-(3-thienyl)-4-methoxybenzaldehyde, 2-methyl-5-isopropyl-4-methoxy-benzaldehyde, 2,3-dihydrobenzofuran-5-carboxaldehyde, 2,2-dimethylchroman-6-carboxaldehyde, 3-(thiophen-3-yl)-4-methoxybenzaldehyde, 3-(3-acetylaminophenyl)-4-methoxy-benzaldehyde, 2-naphthyl-4,5-dimethoxybenzaldehyde, 2-(3-methoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-(pyridin-3-yl)-p-anisaldehyde, 2-(3-ethoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-isopropyl-4-[2-(N-morpholino)ethoxy]benzaldehyde, 3,5-diisopropyl-4-[2-(N-morpholino)ethoxy]benzaldehyde, 2-(2-methoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-(3-ethoxyphenyl)-p-anisaldehyde, 3-(thiophen-2-yl)-4-[2-(N-morpholino)ethoxy]-benzaldehyde, 2-(thiophen-2-yl)-4,5-dimethoxybenzaldehyde, 2,4-dimethyl-3-[3-dimethylaminopropyl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[3-carboxypropyl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[3-(N-morpholino)prop-1-yl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[2-dimethylaminoethyl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[2-carboxyethyl]-pyrrole-5-carboxaldehyde and 2,4-dimethyl-3-[2-(N-morpholino)propyl]-pyrrole-5-carboxaldehyde, 3-isopropyl-4-[2-(N-morpholino)propoxy]benzaldehyde, 3,5-diisopropyl-4-[2-(N-morpholino)propoxy]benzaldehyde, 3-(thiophen-2-yl)-4-[2-N-morpholino)propoxy]benzaldehyde,.
Another aspect of this invention provides a method for the synthesis of 3-arylidenyl-6-heterocyclyl-2-indolinone of formula I comprising condensing an oxindole of formula 2 with an aldehyde of formula 3 in a solvent, preferably in the presence of a base.
Examples of the oxindoles of formula 2 that may be condensed with an aldehyde of formula 3 to give a 3-arylidenyl-6-heterocyclyl-2-indolinones of formula I are 6-(pyridin-2-yl)-2-oxindole, 6-(pyridin-3-yl)-2-oxindole, 6-(pyridine-4-yl)-2-oxindole, 6-(pyrimidin-2-yl)-2-oxindole, 6-(pyrimidin-4-yl)-2-oxindole, 6-(pyrimidin-5-yl)-2-oxindole, 6-(triazinyl)-2-oxindole, 6-(pyrrol-2-yl)-2-oxindole, 6-(pyrrol-3-yl)-2-oxindole, 6-(thiophen-2-yl)-2-oxindole, 6-(thiophen-3-yl)-2-oxindole, 6-(furan-2-yl)-2-oxindole, 6-(furan-3-yl)-2-oxindole, 6-(imidazol-2-yl)-2-oxindole, 6-(imidazol-4-yl)-2-oxindole, 6-(thiazol-2-yl)-2-oxindole, 6-(thiazol-4-yl)-2-oxindole, 6-(oxazol-2-yl)-2-oxindole, 6-(oxazol-4-yl)-2-oxindole, 6-(thiadiazol-2-yl)-2-oxindole, 6-(oxadiazol-2-yl)-2-oxindole and 6-(triazol-2-yl)-2-oxindole, 6-(3methylisoxazole-5-yl)-2-oxindole, 6-(3-methylisothiazole-5-yl)-2-oxindole, 6-(3,5-dimethyl-isoxazole-4-yl)-2-oxindole, 6-(thiazole-2-yl)-2-oxindole, 6-(thiazole-4-yl)-2-oxindole, 6-(thiazole-5-yl)-2-oxindole, 6-(3-methylthiophene-2-yl)-2-oxindole, 6-(4-methylthiophene-2-yl)-2-oxindole, 6-(5-methylthiophene-2-yl)-2-oxindole, 6-(5-chlorothiophene-2-yl)-2-oxindole, 6-(4-methylfuran-2-yl)-2-oxindole.
Examples of aldehydes of structure 3 which may be condensed with oxindoles of structure 2 to give a compound of this invention are, without limitation, benzaldehyde, 3-isopropyl-p-anisaldehyde, 2-methyl-5-isopropyl-p-anisaldehyde, 3,5-diisopropyl-p-anisaldehyde, 3-cyclopentyl-p-anisaldehyde, 3-cyclohexyl-p-anisaldehyde, 3-phenyl-p-anisaldehyde, 3,5-dimethyl-p-anisaldehyde, 2-hydroxy-3,5-dichlorobenzaldehyde, 2-hydroxy-5-chlorobenzaldehyde, 2-hydroxy-4-methoxy-5-(4-methoxyphenyl)benzaldehyde, 3-cyclopentyl-4-methoxybenzaldehyde, 3-cyclopentyl-4-hydroxybenzaldehyde, 3-(2-thienyl)-4-methoxybenzaldehyde, 2-hydroxybenzaldehyde, 2-hydroxy-4-methoxybenzaldehyde, 2-hydroxy-5-bromobenzaldehyde, 2-methylpyridine-6-carboxaldehyde, 3-tert-butyl-4-hydroxy-5-bromobenzaldehyde, 3-bromobenzaldehyde, hydroxy-3-fluorobenzaldehyde, 3-cyclohexyl-4-[2-(N-morpholino)ethyl]benzaldehyde, 3-(3-thienyl)-4-methoxybenzaldehyde, 2-methyl-5-isopropyl-4-methoxy-benzaldehyde, 2,3-dihydrobenzofuran-5-carboxaldehyde, 2,2-dimethylchroman-6-carboxaldehyde, 3-(thiophen-3-yl)-4-methoxybenzaldehyde, 3-(3-acetylaminophenyl)-4-methoxy-benzaldehyde, 2-naphthyl-4,5-dimethoxybenzaldehyde, 2-(3-methoxypheny-4,5-dimethoxybenzaldehyde, 3-(pyridin-3-yl)-p-anisaldehyde, 2-(3-ethoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-isopropyl-4-[2-(N-morpholino)ethoxy]benzaldehyde, 3,5-diisopropyl -4-[2-(N-morpholino)ethoxy]benzaldehyde, 2-(2-methoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-(3-ethoxyphenyl)-p-anisaldehyde, 3-(thiophen-2-yl)-4-[2-(N-morpholino)ethoxy]-benzaldehyde, 2-(thiophen-2-yl)-4,5-dimethoxybenzaldehyde, 2,4-dimethyl-3-[3-dimethylaminopropyl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3[2-carboxypropyl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[3-(N-morpholino)propyl-1-yl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[2-dimethylaminoethyl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[2-carboxyethyl]-pyrrole-5-carboxaldehyde and 2,4-dimethyl-3-[2-(N-morpholino)ethyl]prop-1-yl]-pyrrole-5-carboxaldehyde, 3-isopropyl-4-[2-(N-morpholino)propoxy]benzaldehyde, 3,5-di isopropyl-4-[2-(N-morpholino)propoxy]benzaldehyde, 3-(thiophen-2-yl)-4-[2-N-morpholino)propoxy]benzaldehyde.
Another aspect of this invention is a combinatorial library of at least ten 3-aralkyl-2-indolinone compounds that can be formed by condensing oxindoles of structure 4 with aldehydes of structure 5 and then reducing the 3-position double bond of the resultant 3-arylidene-2-oxindole. In particular, the condensation refers to reaction xe2x80x9cAxe2x80x9d in Scheme II. Compound 4 is the xe2x80x9c3-arylidene-2-oxindolexe2x80x9d referred to above.
xe2x80x9cReducing,xe2x80x9d as used herein, refers to the addition of hydrogen across the double 
(Reaction xe2x80x9cBxe2x80x9d) to give compound II in Scheme II.
The oxindole in the above combinatorial library is preferrably selected from the group consisting of oxindole itself and substituted oxindoles such as, without limitation, 5-fluorooxindole, 6-fluorooxindole, 7-fluorooxindole, 6-trifluoromethyloxindole, 5-chlorooxindole, 6-chlorooxindole, indole-4-carboxylic acid, 5-bromooxindole, 6-(acetamido)-oxindole, 4-methyloxindole, 5-methyloxindole, 4-methyl-5-chlorooxindole, 5-ethyloxindole, 6-hydroxyoxindole, 6-(cyclopentylcarboxamido)oxindole, 5-acetyloxindole, oxindole-5-carboxylic acid, 5-methoxyoxindole, 6-methoxyoxindole, 5-aminooxindole, 6-aminooxindole, 4-[2-(N-morpholino)ethyl]-oxindole, 7-azaoxindole, oxindole-4-carabamic acid t-butyl ester, oxindole-6-carbamic acid t-butyl ester, 4-(2-carboxyethyl)oxindole, 4-n-butyloxindole, 4,5-dimethoxyoxindole, 6-(methanesulfonamido)oxindole, 6-(benzamido)oxindole, 5-ethoxyoxindole, 6-phenyloxindole, 4-(2-hydroxyeth-1-yl)oxindole, 6-(2-methoxyphen-1-yl)oxindole, 6-(3-methoxyphen-1-yl)oxindole and 6-(4-methoxyphen-1-yl)oxindole.
The aldehyde in the above combinatorial library is preferably selected from the group consisting of, without limitation, benzaldehyde itself as well as 3-isopropyl-p-anisaldehyde, 2-methyl-5-isopropyl-p-anisaldehyde, 3,5-diisopropyl-p-anisaldehyde, 3-cyclopentyl-p-anisaldehyde, 3-cyclohexyl-p-anisaldehyde, 3-phenyl-p-anisaldehyde, 3,5-dimethyl-p-anisaldehyde, 2-hydroxy-3,5-dichlorobenzaldehyde, 2-hydroxy-5-chlorobenzaldehyde, 2-hydroxy-4-methoxy-5-(4-methoxyphenyl)benzaldehyde, 3-cyclopentyl-4-methoxybenzaldehyde, 3-cyclopentyl-4-hydroxybenzaldehyde, 3-(2-thienyl)-4-methoxybenzaldehyde, 2-hydroxybenzaldehyde, 2-hydroxy-4-methoxybenzaldehyde, 2-hydroxy-5-bromobenzaldehyde, 2-methylpyridine-6-carboxaldehyde, 3-tert-butyl-4-hydroxy-5-bromobenzaldehyde, 3-bromobenzaldehyde, 3,5-diisopropyl-4-[2-(N-morpholino)ethyl]benzaldehyde,indole-5-carboxaldehyde, 2-hydroxy-3-fluorobenzaldehyde, 3-cyclohexyl-4-[2-(N-morpholino)ethyl]benzaldehyde, 3-(3-thienyl)-4-methoxybenzaldehyde, 2-methyl-5-isopropyl-4-methoxy-benzaldehyde, 2,3-dihydrobenzofuran-5-carboxaldehyde, 2,2-dimethylchroman-6-carboxaldehyde, 3-(thiophen-3-yl)-4-methoxybenzaldehyde, 3-(3-acetylaminophenyl)-4-methoxy-benzaldehyde, 2-naphthyl-4,5-dimethoxybenzaldehyde, 2-(3-methoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-(pyrid-3-yl)-p-anisaldehyde, 2-(3-ethoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-isopropyl-4-[2-(N-morpholino)ethoxy]benzaldehyde, 3,5-diisopropyl-4-[2-(N-morpholino)ethoxy]benzaldehyde, 2-(2-methoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-(3-ethoxyphenyl)-p-anisaldehyde, 3-(thiophen-2-yl)-4-[2-(N-morpholino)ethoxy]-benzaldehyde, 2-(thiophen-2-yl)-4,5-dimethoxybenzaldehyde, 2,4-dimethyl-3-[3-dimethylaminoprop-1-yl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[3-carboxyprop-1-yl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[3-(N-morpholino)prop-1-yl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[2-dimethylaminoethyl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[2-carboxyethyl]-pyrrole-5-carboxaldehyde and 2,4-dimethyl-3-[2-(N-morpholino)ethyl]prop-1-yl]-pyrrole-5-carboxaldehyde.
Another aspect of this invention provides a method for the synthesis of 3-aralkyl-2-indolinone of formula II comprising condensing an oxindole of formula 4 with an aldehyde of formula 5 in a solvent, preferably in the presence of a base, optionally isolating the resultant 3-arylidene-2-oxindole and then reducing the 3-arylidene-2-oxindole.
Examples of the oxindoles of formula 4 that may be condensed with an aldehyde of formula 5 and the product reduced to give the 3-aralkyl-2-indolinones of formula II are oxindole itself and substituted oxindoles such as, without limitation, 5-fluorooxindole, 6-fluorooxindole, 7-fluorooxindole, 6-trifluoromethyloxindole, 5-chlorooxindole, 6-chlorooxindole, indole-4-carboxylic acid, 5-bromooxindole, 6-(acetamido)-oxindole, 4-methyloxindole, 5-methyloxindole, 4-methyl-5-chlorooxindole, 5-ethyloxindole, 6-hydroxyoxindole, 6-(cyclopentylcarboxamido)oxindole, 5-acetyloxindole, oxindole-5-carboxylic acid, 5-methoxyoxindole, 6-methoxyoxindole, 5-aminooxindole, 6-aminooxindole, 4-[2-(N-morpholino)ethyl]-oxindole, 7-azaoxindole, oxindole-4-carabamic acid t-butyl ester, oxindole-6-carbamic acid t-butyl ester, 4-(2-carboxyethyl)oxindole, 4-n-butyloxindole, 4,5-dimethoxyoxindole, 6-(methanesulfonamido)oxindole, 6-(benzamido)oxindole, 5-ethoxyoxindole, 6-phenyloxindole, 4-(2-hydroxyeth-1-yl)oxindole, 6-(2-methoxyphen-1-yl)oxindole, 6-(3-methoxyphen-1-yl)oxindole and 6-(4-methoxyphen-1-yl)oxindole.
Examples of aldehydes of structure 3 which may be condensed with oxindoles of structure 4 and the product reduced to give a compound of this invention are, without limitation, benzaldehyde itself as well as 3-isopropyl-p-anisaldehyde, 2-methyl-5-isopropyl-p-anisaldehyde, 3,5-diisopropyl-p-anisaldehyde, 3-cyclopentyl-p-anisaldehyde, 3-cyclohexyl-p-anisaldehyde, 3-phenyl-p-anisaldehyde, 3,5-dimethyl-p-anisaldehyde, 2-hydroxy-3,5-dichlorobenzaldehyde, 2-hydroxy-5-chlorobenzaldehyde, 2-hydroxy-4-methoxy-5-(4-methoxyphenyl)benzaldehyde, 3-cyclopentyl-4-methoxybenzaldehyde, 3-cyclopentyl-4-hydroxybenzaldehyde, 3-(2-thienyl)-4-methoxybenzaldehyde, 2-hydroxybenzaldehyde, 2-hydroxy-4-methoxybenzaldehyde, 2-hydroxy-5-bromobenzaldehyde, 2-methylpyridine-6-carboxaldehyde, 3-tert-butyl-4-hydroxy-5-bromobenzaldehyde, 3-bromobenzaldehyde, 3,5-diisopropyl-4-[2-(N-morpholino)ethyl]benzaldehyde,indole-5-carboxaldehyde, 2-hydroxy-3-fluorobenzaldehyde, 3-cyclohexyl-4-[2-(N-morpholino)ethyl]benzaldehyde, 3-(3-thienyl)-4-methoxybenzaldehyde, 2-methyl-5-isopropyl-4-methoxybenzaldehyde, 2,3-dihydrobenzofuran-5-carboxaldehyde, 2,2-dimethylchroman-6-carboxaldehyde, 3-(thiophen-3-yl)-4-methoxybenzaldehyde, 3-(3-acetylaminophenyl)-4-methoxybenzaldehyde, 2-naphthyl-4,5-dimethoxybenzaldehyde, 2-(3-methoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-(pyrid-3-yl)-p-anisaldehyde, 2-(3-ethoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-isopropyl-4-[2-(N-morpholino)ethoxy]benzaldehyde, 3,5-diisopropyl-4-[2-(N-morpholino)ethoxy]benzaldehyde, 2-(2-methoxyphenyl)-4,5-dimethoxybenzaldehyde, 3-(3-ethoxyphenyl)-p-anisaldehyde, 3-(thiophen-2-yl)-4-[2-(N-morpholino)ethoxy]-benzaldehyde, 2-(thiophen-2-yl)-4,5-dimethoxybenzaldehyde, 2,4-dimethyl-3-[3-dimethylaminoprop-1-yl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[3-carboxyprop-1-yl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[3-(N-morpholino)prop-1-yl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[2-dimethylaminoethyl]-pyrrole-5-carboxaldehyde, 2,4-dimethyl-3-[2-carboxyethyl]-pyrrole-5-carboxaldehyde and 2,4-dimethyl-3-[2-(N-morpholino)ethyl]prop-1-yl]-pyrrole-5-carboxaldehyde.
In another aspect, the invention provides a combinatorial library of at least 10 indolinone compounds that can be formed by reacting an oxindole with a ketone, where the oxindole has the following structure 
and the ketone has the following structure 
and R27, R28, R29, R30, R31, R32, R33, and R34 are as described herin.
The oxindole in the above combinatorial library is preferrably selected from the group consisting of 1,3-dihydro-indol-2-one, 4-methyl-1,3-dihydro-indol-2-one, 4-(2-hydroxy-ethyl)-1,3-dihydro-indol-2-one, 5-chloro-1,3-dihydro-indol-2-one, 5-bromo-1,3-dihydro-indol-2-one, 5-methoxy-1,3-dihydro-indol-2-one, 2-oxo-2,3-dihydro-1H-indole-5-carboxylic acid, 2-oxo-2,3-dihydro-1H-indole-5-sulfonic acid dimethylamide, N-(2-oxo-2,3-dihydro-1H-indole-5-yl)-acetamide, 6-chloro-1,3-dihydro-indol-2-one, 6-methoxy-1,3-dihydro-indol-2-one, and N-(5-methyl-2-oxo-2,3-dihydro-1H-indole-6-yl)-acetamide.
The ketone is preferably selected from the group consisting of 
Another aspect of the invention provides for a method for synthesizing a compound of formula III, as described herein, comprising the step of reacting a first reactant with a second reactant in a solvent and in the presence of a base at elevated temperatures, where the first reactant is an oxindole having the following structure 
and the second reactant is a ketone having the following structure 
and R27, R28, R29, R30, R31, R32, R33, and R34 are as described herin.
The first reactant is preferrably an oxindole selected from the group consisting of 1,3-dihydro-indol-2-one, 4-methyl-1,3-dihydro-indol-2-one, 4-(2-hydroxy-ethyl)-1,3-dihydro-indol-2-one, 5-chloro-1,3-dihydro-indol-2-one, 5-bromo-1,3-dihydro-indol-2-one, 5-methoxy-1,3-dihydro-indol-2-one, 5-carboxy-1,3-dihydro-indol-2-one, 5-dimethylsulfonamido-1,3-dihydro-indol-2-one, 5-formamido-1,3-dihydro-indol-2-one, 6-chloro-1,3-dihydro-indol-2-one, 6-methoxy-1,3-dihydro-indol-2-one, and 5-methyl-6-formamido-1,3-dihydro-indol-2-one.
The second reactant is preferrably a ketone selected from the group consisting of 
In another aspect, the invention provides a combinatorial library of at least 10 indolinone compounds that can be formed by reacting an oxindole with an aldehyde, where the oxindole has a structure set forth in formula I, as defined herein, and where the aldehyde has the formula
Zxe2x80x94C(O)xe2x80x94H
where Z is as defined herein.
The oxindole in the above combinatorial library is preferrably selected from the group consisting of 4-[2-(3-isopropyl-phenoxy)-ethyl]-1,3-dihydro-indol-2-one, 4-[2-(2-isopropyl-phenoxy)-ethyl]-1,3-dihydro-indol-2-one, 4-[2-(biphenyl-3-yloxy)-ethyl]-1,3-dihydro-indol-2-one, 4-(2-hydroxy-ethyl)-1,3-dihydro-indol-2-one, phenyl-thiocarbamic acid O-[2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl]ester, phenyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, tert-butyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, cyclohexyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, benzene sulfonyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, biphenyl-2-yl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, ethyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, 4-[2-(4-methoxy-phenoxy)-ethyl]-1,3-dihydro-indol-2-one, 4-(2-methoxy-ethyl)-1,3-dihydro-indol-2-one, 4-(2-ethoxy-ethyl)-1,3-dihydro-indol-2-one, 4-[2-(4-isopropyl-phenoxy)-ethyl]-1,3-dihydro-indol-2-one, 4-[2-(5-chloro-pyridin-3-yloxy)-ethyl]-1,3-dihydro-indol-2-one, and isopropyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester.
The aldehyde is preferably selected from the group consisting of 
Another aspect of the invention provides for a method for synthesizing a compound of formula IV or formula V, as described herein. The method of synthesizing a compound of formula IV, comprises the step of reacting a first reactant with a second reactant in a solvent and in the presence of a mixture of other reagents. The reaction may take place at ambient temperatures or at elevated temperatures.
The first reactant is preferably an oxindole, more preferably 4-(2-hydroxyethyl)-1,3-dihydro-indol-2-one. The second reactant is preferably selected from the group consisting of an alcohol, an iodoalkyl, an alkyl or aryl isocyanate, and an alkyl or aryl isothiocyanate, and more preferably is selected from the group consisting of 3-isopropylphenol, 2-isopropylphenol, 3-phenylphenol, 4-methoxyphenol, 5-chloro-3-pyridinol, methyl iodide, ethyl iodide, phenyl isocyanate, tert-butyl isocyanate, cyclohexyl isocyanate, benzenesulfonyl isocyanate, 2-biphenylyl isocyanate, ethyl isocyanate, isopropyl isocyanate, and phenyl isothiocyanate.
The xe2x80x9cmixture of other reagentsxe2x80x9d generally refers to a mixture of synthetic reagents or solvents that would facilitate the synthesis of the compounds of the invention, and may include a mixture of diethyl azodicarboxylate with triphenylphosphine in a solvent, or silver trifluoromethanesulfonate, by itself or with a base, in a solvent, or a mixture of solvents.
The method of synthesizing a compound of formula V, comprises the step of reacting a first reactant with a second reactant in a solvent and in the presence of a base at elevated temperatures, where the first reactant is an oxindole having a structure set forth in formula I, as defined herein, and the second reactant is an aldehyde, having the formula
Zxe2x80x94C(O)xe2x80x94H
where Z is as defined herein.
The first reactant is preferrably an oxindole selected from the group consisting of 4-[2-(3-isopropyl-phenoxy)-ethyl]-1,3-dihydro-indol-2-one, 4-[2-(2-isopropyl-phenoxy)-ethyl]-1,3-dihydro-indol-2-one, 4-[2-(biphenyl-3-yloxy)-ethyl]-1,3-dihydro-indol-2-one, 4-(2-hydroxy-ethyl)-1,3-dihydro-indol-2-one, phenyl-thiocarbamic acid O-[2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl]ester, phenyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, tert-butyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, cyclohexyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, benzene sulfonyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, biphenyl-2-yl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, ethyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester, 4-[2-(4-methoxy-phenoxy)-ethyl]-1,3-dihydro-indol-2-one, 4-(2-methoxy-ethyl)-1,3-dihydro-indol-2-one, 4-(2-ethoxy-ethyl)-1,3-dihydro-indol-2-one, 4-[2-(4-isopropyl-phenoxy)-ethyl]-1,3-dihydro-indol-2-one, 4-[2-(5-chloro-pyridin-3-yloxy)-ethyl]-1,3-dihydro-indol-2-one, and isopropyl-carbamic acid 2-(2-oxo-2,3-dihydro-1H-indol-4-yl)-ethyl ester.
The second reactant is preferrably an aldehyde selected from the group consisting of 4-bromobenzaldehyde, 2-pyridinecarboxaldehyde, 6-methyl-2-pyridinecarbaldehyde, 1H-indole-5-carbaldehyde, 4-methoxy-3-thiophen-2-yl-benzaldehyde, 4-(3-dimethylamino-propyl)-3,5-dimethyl-1H-pyrrole-2-carbaldehyde, 3-hydroxy-6-ethyl-pyridine-2-carbaldehyde, 4-(3-dimethylamino-propyl)-3,5-dimethyl-1H-pyrrole-2-carbaldehyde, and 4-carboxyethyl-3,5-dimethyl-2-formylpyrrole.
In another aspect, the invention features a pharmaceutical composition comprising (i) a physiologically acceptable carrier, diluent, or excipient; and (ii) a compound as described herein.
The invention also features a method of modulating the function of a protein kinase with a compound of the invention, comprising the step of contacting cells expressing the protein kinase with the compound.
A still further aspect of this invention is that the protein kinase whose catalytic activity is being modulated by a compound of this invention is selected from the group consisting of receptor protein tyrosine kinases, cellular tyrosine kinases and serine-threonine kinases.
It is an aspect of this invention that the receptor protein kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of EGF, HER2, HER3, HER4, IR, IGF-1R, IRR, PDGFRxcex1, PDGFRxcex2, CSFIR, C-Kit, C-fms, Flk-1R, Flk4, KDR/Flk-1, Flt-1, FGFR-1R, FGFR-2R, FGFR-3R and FGFR-4R. In addition, it is an aspect of this invention that the cellular tyrosine kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of Src, Frk, Btk, Csk, Abl, ZAP70, Fes/Fps, Fak, Jak, Ack, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk. Another aspect of this invention is that the serine-threonine protein kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of CDK2 and Raf.
A protein kinase natural binding partner can bind to a protein kinase""s intracellular region with high affinity. High affinity represents an equilibrium binding constant on the order of 10xe2x88x926 M or less. In addition, a natural binding partner can also transiently interact with a protein kinase intracellular region and chemically modify it. Protein kinase natural binding partners are chosen from a group that includes, but is not limited to, SRC homology 2 (SH2) or 3 (SH3) domains, other phosphoryl tyrosine binding (PTB) domains, guanine nucleotide exchange factors, protein phosphatases, and other protein kinases. Methods of determining changes in interactions between protein kinases and their natural binding partners are readily available in the art.
The compounds of the invention preferably modulate the activity of the protein tyrosine kinase in vitro. These compounds preferably show positive results in one or more in vitro assays for an activity corresponding to treatment of the disease or disorder in question (such as the assays described in the Examples below).
The invention also features a method of identifying compounds that modulate the function of protein kinase, comprising the following steps: (a) contacting cells expressing the protein tyrosine kinase with the compound; and (b) monitoring an effect upon the cells. The effect upon the cells is preferably a change or an absence of a change in cell phenotype, more preferably it is a change or an absence of a change in cell proliferation, even more preferably it is a change or absence of a change in the catalytic activity of the protein kinase, and most preferably it is a change or absence of a change in the interaction between the protein kinase with a natural binding partner, as described herein.
In a preferred embodiment, the invention features a method for identifying the compounds of the invention, comprising the following steps: (a) lysing the cells to render a lysate comprising protein tyrosine kinase; (b) adsorbing the protein tyrosine kinase to an antibody; (c)incubating the adsorbed protein tyrosine kinase with a substrate or substrates; and (d) adsorbing the substrate or substrates to a solid support or antibody; where the step of monitoring the effect on the cells comprises measuring the phosphate concentration of the substrate or substrates.
In yet another aspect, the invention features a method for treating a disease related to unregulated kinase signal transduction, where the method includes the step of administering to a subject in need thereof a therapeutically effective amount of a compound of the invention as described herein.
The invention also features a method of regulating kinase signal transduction comprising administering to a subject a therapeutically effective amount of a compound of the invention as described herein.
Furthermore, the invention features a method of preventing or treating an abnormal condition in an organism, where the abnormal condition is associated with an aberration in a signal transduction pathway characterized by an interaction between a protein kinase and a natural binding partner, where the method comprises the following steps: (a) administering a compound of the invention as described herein; and (b) promoting or disrupting the abnormal interaction. The organism is preferably a mammal and the abnormal condition is preferably cancer. The abnormal condition may also preferably be selected from the group consisting of hypertension, depression, generalized anxiety disorder, phobias, post-traumatic stress syndrome, avoidant personality disorder, sexual dysfunction, eating disorders, obesity, chemical dependencies, cluster headache, migraine, pain, Alzheimer""s disease, obsessive-compulsive disorder, panic disorder, memory disorders, Parkinson""s disease, endocrine disorders, vasospasm, cerebellar ataxia, and gastrointestinal tract disorders.
As used herein, xe2x80x9cPK related disorder,xe2x80x9d xe2x80x9cPK driven disorder,xe2x80x9d and xe2x80x9cabnormal PK activityxe2x80x9d all refer to a condition characterized by inappropriate; i.e., under or, more commonly, over, PK catalytic activity, where the particular PK can be an RTK, a CTK or an STK. Inappropriate catalytic activity can arise as the result of either: (1) PK expression in cells which normally do not express PKs; (2) increased PK expression leading to unwanted cell proliferation, differentiation and/or growth; or, (3) decreased PK expression leading to unwanted reductions in cell proliferation, differentiation and/or growth. Over-activity of a PK refers to either amplification of the gene encoding a particular PK or production of a level of PK activity which can correlate with a cell proliferation, differentiation and/or growth disorder (that is, as the level of the PK increases, the severity of one or more of the symptoms of the cellular disorder increases). Under-activity is, of course, the converse, wherein the severity of one or more symptoms of a cellular disorder increase as the level of the PK activity decreases.
The term xe2x80x9ctherapeutically effective amountxe2x80x9d as used herein refers to that amount of the compound being administered which will relieve to some extent one or more of the symptoms of the disorder being treated. In reference to the treatment of cancer, a therapeutically effective amount refers to that amount which has the effect of (1) reducing the size of the tumor; (2) inhibiting (that is, slowing to some extent, preferably stopping) tumor metastasis; (3) inhibiting to some extent (that is, slowing to some extent, preferably stopping) tumor growth; and/or, (4) relieving to some extent (or, preferably, eliminating) one or more symptoms associated with the cancer.
It is an aspect of this invention that the above-referenced protein kinase related disorder is selected from the group consisting of a receptor protein tyrosine kinase related disorder, a cellular tyrosine kinase disorder and a serine-threonine kinase related disorder.
In yet another aspect of this invention, the above referenced protein kinase related disorder is selected from the group consisting of an EGFR related disorder, a PDGFR related disorder, an IGFR related disorder and a flk related disorder.
The above referenced protein kinase related disorder is a cancer selected from the group consisting of squamous cell carcinoma, astrocytoma, glioblastoma, lung cancer, bladder cancer, head and neck cancer, melanoma, ovarian cancer, prostate cancer, breast cancer, small-cell lung cancer and glioma in a further aspect of this invention.
The above referenced protein kinase related disorder is selected from the group consisting of diabetes, immunlogical disorders such as autoimmune disorder, a hyperproliferation disorder, restinosis, fibrosis, psoriasis, osteoarthritis, rheumatoid arthritis, an inflammatory disorder, angiogenesis, cardiovascular disease such as aetherosclerosis, and renal disease, in yet another aspect of this invention.
In addition to modulating PK activity, the compounds of this invention may inhibit the activity of protein phosphatases which are enzymes which remove phosphate groups from phosphorylated proteins. Thus the compounds disclosed herein may also represent a new generation of therapeutic compounds for diseases and disorders associated with abnormal phosphatase activity (such as, without limitation, diabetes, cell proliferation disorders and inflammatory disorders). The terms defined herein with respect to PKs would be understood by one skilled in the art to have the same or similar meaning with regard to phosphastases.
The summary of the invention described above is non-limiting and other features and advantages of the invention will be apparent from the following description of the preferred embodiments, and from the claims.
The present invention relates to compounds capable of regulating and/or modulating cellular signal transduction and, in preferred embodiments, receptor and non-receptor tyrosine kinase signal transduction.
Receptor-kinase mediated signal transduction is initiated by extracellular interaction with a specific growth factor (ligand), followed by receptor dimerization, transient stimulation of the intrinsic protein kinase activity and phosphorylation. Binding sites are thereby created for intracellular signal transduction molecules and lead to the formation of complexes with a spectrum of cytoplasmic signaling molecules that facilitate the appropriate cellular response (e.g., cell division, metabolic effects to the extracellular microenvironment). See, Schlessinger and Ullrich, 1992, Neuron 9:303-391.
It has been shown that tyrosine phosphorylation sites in growth factor receptors unction as high-affinity binding sites for SH2 (src homology) domains of signaling molecules. Fantl et al., 1992, Cell 69:413-423; Songyang et al., 1994, Mol. Cell. Biol. 4:2777-2785); Songyang et al., 1993, Cell 72:767-778; and Koch et al., 1991, Science 252:668-678. Several intracellular substrate proteins that associate with receptor kinases have been identified. They may be divided into two principal groups: (1) substrates which have a catalytic domain; and (2) substrates which lack such domain but serve as adapters and associate with catalytically active molecules. Songyang et al., 1993, Cell 72:767-778. The specificity of the interactions between receptors and SH2 domains of their substrates is determined by the amino acid residues immediately surrounding the phosphorylated tyrosine residue. Differences in the binding affinities between SH2 domains and the amino acid sequences surrounding the phosphotyrosine residues on particular receptors are consistent with the observed differences in their substrate phosphorylation profiles. Songyang et al., 1993, Cell 72:767-778. These observations suggest that the function of each receptor kinase is determined not only by its pattern of expression and ligand availability but also by the array of downstream signal transduction pathways that are activated by a particular receptor. Thus, phosphorylation provides an important regulatory step which determines the selectivity of signaling pathways recruited by specific growth factor receptors, as well as differentiation factor receptors.
Kinase signal transduction results in, among other responses, cell proliferation, differentiation and metabolism. Abnormal cell proliferation may result in a wide array of disorders and diseases, including the development of neoplasia such as carcinoma, sarcoma, leukemia, glioblastoma, hemangioma, psoriasis, arteriosclerosis, arthritis and diabetic retinopathy (or other disorders related to uncontrolled angiogenesis and/or vasculogenesis).
This invention is therefore directed to compounds which regulate, modulate and/or inhibit kinase signal transduction by affecting the enzymatic activity of the receptor kinases (RKs) and/or the non-receptor kinases and interfering with the signal transduced by such proteins. More particularly, the present invention is directed to compounds which regulate, modulate and/or inhibit the RK and/or non-receptor kinase mediated signal transduction pathways as a therapeutic approach to cure many kinds of solid tumors, including but not limited to carcinoma, sarcoma, erythroblastoma, glioblastoma, meningioma, astrocytoma, melanoma and myoblastoma. Indications may include, but are not limited to brain cancers, bladder cancers, ovarian cancers, gastric cancers, pancreas cancers, colon cancers, blood cancers, lung cancers and bone cancers.
The compounds described herein are useful for treating disorders related to unregulated kinase signal transduction, including cell proliferative disorders, fibrotic disorders-and metabolic disorders.
Cell proliferative disorders which can be treated or further studied by the present invention include cancers, blood vessel proliferative disorders and mesangial cell proliferative disorders.
Blood vessel proliferative disorders refer to angiogenic and vasculogenic disorders generally resulting in abnormal proliferation of blood vessels. The formation and spreading of blood vessels, or vasculogenesis and angiogenesis, respectively, play important roles in a variety of physiological processes such as embryonic development, corpus luteum formation, wound healing and organ regeneration. They also play a pivotal role in cancer development. Other examples of blood vessel proliferation disorders include arthritis, where new capillary blood vessels invade the joint and destroy cartilage, and ocular diseases, like diabetic retinopathy, where new capillaries in the retina invade the vitreous, bleed and cause blindness. Conversely, disorders related to the shrinkage, contraction or closing of blood vessels, such as restenosis, are also implicated.
Fibrotic disorders refer to the abnormal formation of extracellular matrix. Examples of fibrotic disorders include hepatic cirrhosis and mesangial cell proliferative disorders. Hepatic cirrhosis is characterized by the increase in extracellular matrix constituents resulting in the formation of a hepatic scar. Hepatic cirrhosis can cause diseases such as cirrhosis of the liver. An increased extracellular matrix resulting in a hepatic scar can also be caused by viral infection such as hepatitis. Lipocytes appear to play a major role in hepatic cirrhosis. Other fibrotic disorders implicated include atherosclerosis (see, below).
Mesangial cell proliferative disorders refer to disorders brought about by abnormal proliferation of mesangial cells. Mesangial proliferative disorders include various human renal diseases, such as glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis, thrombotic microangiopathy syndromes, transplant rejection, and glomerulopathies. The PDGF-R has been implicated in the maintenance of mesangial cell proliferation. Floege et al., 1993, Kidney International 43:47S-54S.
PKs have been associated with such cell proliferative disorders. For example, some members of the receptor tyrosine kinase family have been associated with the development of cancer. Some of these receptors, like the EGFR (Tuzi et al., 1991, Br. J. Cancer 63:227-233; Torp et al., 1992, APMIS 100:713-719) HER2/neu (Slamon et al., 1989, Science 244:707-712) and the PDGF-R (Kumabe et al., 1992, Oncogene 7:627-633) are overexpressed in many tumors and/or persistently activated by autocrine loops. In fact, in the most common and severe cancers these receptor overexpressions (Akbasak and Suner-Akbasak et al., 1992, J. Neurol. Sci. 111:119-133; Dickson et al., 1992, Cancer Treatment Res. 61:249-273; Korc et al., 1992, J. Clin. Invest. 90:1352-1360) and autocrine loops (Lee and Donoghue, 1992, J. Cell. Biol. 118:1057-1070; Korc et al., supra; Akbasak and Suner-Akbasak et al., supra) have been demonstrated. For example, the EGFR receptor has been associated with squamous cell carcinoma, astrocytoma, glioblastoma, head and neck cancer, lung cancer and bladder cancer. HER2 has been associated with breast, ovarian, gastric, lung, pancreas and bladder cancer. The PDGF-R has been associated with glioblastoma, lung, ovarian, melanoma and prostate cancer. The RK c-met has been generally associated with hepatocarcinogenesis and thus hepatocellular carcinoma. Additionally, c-met has been linked to malignant tumor formation. More specifically, the RK c-met has been associated with, among other cancers, colorectal, thyroid, pancreatic and gastric carcinoma, leukemia and lymphoma. Additionally, over-expression of the c-met gene has been detected in patients with Hodgkin""s disease, Burkitt""s disease, and the lymphoma cell line. Fik has likewise been associated with a broad spectrum of tumors including, without limitation, mammary, ovarian and lung tumors as well as gliomas such as glioblastoma.
The IGF-IR, in addition to being implicated in nutritional support and in type-II diabetes, has also been associated with several types of cancers. For example, IGF-I has been implicated as an autocrine growth stimulator for several tumor types, e.g., human breast cancer carcinoma cells (Arteaga et al., 1989, J. Clin. Invest. 84:1418-1423) and small lung tumor cells (Macauley et al., 1990, Cancer Res. 50:2511-2517). In addition, IGF-I, integrally involved in the normal growth and differentiation of the nervous system, appears to be an autocrine stimulator of human gliomas. Sandberg-Nordqvist et al., 1993, Cancer Res. 53:2475-2478. The importance of the IGF-IR and its ligands in cell proliferation is further supported by the fact that many cell types in culture (fibroblasts, epithelial cells, smooth muscle cells, T-lymphocytes, myeloid cells, chondrocytes, osteoblasts, the stem cells of the bone marrow) are stimulated to grow by IGF-I. Goldring and Goldring, 1991, Eukaryotic Gene Expression 1:301-326. In a series of recent publications, Baserga even suggests that IGF-I-R plays a central role in the mechanisms of transformation and, as such, could be a preferred target for therapeutic interventions for a broad spectrum of human malignancies. Baserga, 1995, Cancer Res. 55:249-252; Baserga, 1994, Cell 79:927-930; Coppola et al., 1994, Mol. Cell. Biol. 14:4588-4595.
Some protein kinases (PKs) have been implicated in many types of cancer including, notably, breast cancer (Cance, et al., Int. J. Cancer, 54:571-77 (1993)).
The association between abnormalities in RKs and disease are not restricted to cancer, however. For example, RKs have been associated with metabolic diseases like psoriasis, diabetes mellitus, wound healing, inflammation, and neurodegenerative diseases. These diseases include, but are not limited to hypertension, depression, generalized anxiety disorder, phobias, post-traumatic stress syndrome, avoidant personality disorder, sexual dysfunction, eating disorders, obesity, chemical dependencies, cluster headache, migraine, pain, Alzheimer""s disease, obsessive-compulsive disorder, panic disorder, memory disorders, Parkinson""s disease, endocrine disorders, vasospasm, cerebellar ataxia, and gastrointestinal tract disorders. For example, the EGF-R is indicated in corneal and dermal wound healing. Defects in the Insulin-R and the IGF-1R are indicated in type-II diabetes mellitus. A more complete correlation between specific RKs and their therapeutic indications is set forth in Plowman et al., 1994, DNandP 7:334-339.
Not only receptor type kinases, but also many cellular kinases (CKs) including src, abl, fps, yes, fyn, lyn, lck, blk, hck, fgr, yrk (reviewed by Bolen et al., 1992, FASEB J. 6:3403-3409) are involved in the proliferative and metabolic signal transduction pathway and thus in indications of the present invention. For example, mutated src (v-src) has been demonstrated as an oncoprotein (pp60v-src) in chicken. Moreover, its cellular homolog, the proto-oncogene pp60c-src transmits oncogenic signals of many receptors. For example, overexpression of EGF-R or HER2/neu in tumors leads to the constitutive activation of pp60cxe2x80x94src which is characteristic for the malignant cell but absent from the normal cell. On the other hand, mice deficient for the expression of c-src exhibit an osteopetrotic phenotype, indicating a key participation of c-src in osteoclast function and a possible involvement in related disorders. Similarly, Zap 70 is implicated in T-cell signaling.
Furthermore, the identification of CTK modulating compounds to augment or even synergize with RK aimed blockers is an aspect of the present invention.
Additionally, both RKs and non-receptor type kinases have been connected to hyperimmune disorders.
Further, the compounds of the present invention are also effective in treating diseases that are related to the PYK-2 protein. This protein, its cellular function, and diseases related to them are set forth in detail in U.S. applications Ser. No. 08/357,642, filed Dec. 15, 1994, by Lev et al., and entitled xe2x80x9cPYK2 RELATED PRODUCTS AND METHODSxe2x80x9d, and Ser. No. 08/460,626, filed Jun. 2, 1995, by Lev et al., and entitled xe2x80x9cPYK2 RELATED PRODUCTS AND METHODSxe2x80x9d, both of which are hereby incorporated by reference herein in their entirety, including any drawings.
Finally, the present invention is directed towards oxindole and indolinone compounds and methods of modulating the functions of protein phosphatases, as well as methods of preventing and treating protein phosphatase related abnormal conditions in organisms with a compound of the method identified above. The compounds of the invention, as well as compounds obtained by adding chemical substituents, may potently inhibit the action of phosphatases and may represent a new generation of therapeutics for diseases associated with defects in said phosphatases. Terms defined above with respect to kinases have a similar meaning to one skilled in the art with respect to phosphatases. Both RKs and non-receptor type kinases have been connected to hyperimmune disorders.
The compounds of the present invention are also effective in treating diseases that are related to the PYK-2 protein. This protein, its cellular function, and diseases related to them are set forth in detail in U.S. Pat. No. 5,837,524, issued Nov. 17, 1998, to Lev et al., and entitled xe2x80x9cPYK2 RELATED PRODUCTS AND METHODS,xe2x80x9d and U.S. Pat. No. 5,837,815, issued Nov. 17, 1998, to Lev et al., and entitled xe2x80x9cPYK2 RELATED PRODUCTS AND METHODS,xe2x80x9d both of which are hereby incorporated by reference herein in their entirety, including any drawings.
a. The KDR/FLK-1 Receptor and VEGF
Normal vasculogenesis and angiogenesis play important roles in a variety of physiological processes such as embryonic development, wound healing, organ regeneration and female reproductive processes such as follicle development in the corpus luteum during ovulation and placental growth after pregnancy. Folkman and Shing, 1992, J. Biological Chem. 267:10931-34. However, many diseases are driven by persistent unregulated or inappropriate angiogenesis. For example, in arthritis, new capillary blood vessels invade the joint and destroy the cartilage. In diabetes, new capillaries in the retina invade the vitreous, bleed and cause blindness. Folkman, 1987, in: Congress of Thrombosis and Haemostasis (Verstraete, et. al, eds.), Leuven University Press, Leuven, pp.583-596. Ocular neovascularization is the most common cause of blindness and dominates approximately twenty (20) eye diseases.
Moreover, vasculogenesis and/or angiogenesis have been associated with the growth of malignant solid tumors and metastasis. A tumor must continuously stimulate the growth of new capillary blood vessels for the tumor itself to grow. Furthermore, the new blood vessels embedded in a tumor provide a gateway for tumor cells to enter the circulation and to metastasize to distant sites in the body. Folkman, 1990, J. Natl. Cancer Inst. 82:4-6; Klagsbrunn and Soker, 1993, Current Biology 3:699-702; Folkman, 1991, J. Natl., Cancer Inst. 82:4-6; Weidner et al., 1991, New Engl. J. Med. 324:1-5.
Several polypeptides with in vitro endothelial cell growth promoting activity have been identified. Examples include acidic and basic fibroblastic growth factor (aFGF, bFGF), vascular endothelial growth factor (VEGF) and placental growth factor. Unlike aFGF and bFGF, VEGF has recently been reported to be an endothelial cell specific mitogen. Ferrara and Henzel, 1989, Biochem. Biophys. Res. Comm. 161:851-858; Vaisman et al., 1990, J. Biol. Chem. 265:19461-19566.
Thus, the identification of the specific receptors to which VEGF binds is an important advancement in the understanding of the regulation of endothelial cell proliferation. Two structurally closely related RKs have been identified to bind VEGF with high affinity: the flt-1 receptor (Shibuya et al., 1990, Oncogene 5:519-524; De Vries et al., 1992, Science 255:989-991) and the KDR/FLK-1 receptor, discussed in the U.S. patent application Ser. No. 08/193,829. Consequently, it had been surmised that these RKs may have a role in the modulation and regulation of endothelial cell proliferation.
Evidence, such as the disclosure set forth in copending U.S. application Ser. No. 08/193,829, strongly suggests that VEGF is not only responsible for endothelial cell proliferation, but also is a prime regulator of normal and pathological angiogenesis. See generally, Klagsbum and Soker, 1993, Current Biology 3:699-702; Houck et al, 1992, J. Biol. Chem. 267:26031-26037. Moreover, it has been shown that KDR/FLK-1 and flt-1 are abundantly expressed in the proliferating endothelial cells of a growing tumor, but not in the surrounding quiescent endothelial cells. Plate et al., 1992, Nature 359:845-848; Shweiki et al., 1992, Nature 359:843-845.
b. Identification Of Agonists And Antagonists To The KDR/FLK-1 Receptor
In view of the deduced importance of RKs in the control, regulation and modulation of endothelial cell proliferation and potentially vasculogenesis and/or angiogenesis, many attempts have been made to identify RK xe2x80x9cinhibitorsxe2x80x9d using a variety of approaches. These include the use of mutant ligands (U.S. Pat. No. 4,966,849); soluble receptors and antibodies (Application No. WO 94/10202; Kendall and Thomas, 1994, Proc. Natl. Acad. Sci. USA 90:10705-10709; Kim et al., 1993, Nature 362:841-844); and RNA ligands (Jellinek et al., 1994, Biochemistry 33:10450-10456).
Furthermore, kinase inhibitors (WO 94/03427; WO 92/21660; WO 91/15495; WO 94/14808; U.S. Pat. No. 5,330,992; Mariani et al., 1994, Proc. Am. Assoc. Cancer Res. 35:2268), and inhibitors acting on receptor kinase signal transduction pathways, such as protein kinase C inhibitors have been identified (Schuchter et al., 1991, Cancer Res. 51:682-687); Takano et al., 1993, Mol. Bio. Cell 4:358A; Kinsella et al., 1992, Exp. Cell Res. 199:56-62; Wright et al., 1992, J. Cellular Phys. 152:448-57).
More recently, attempts have been made to identify small molecules which act as kinase inhibitors for use in the treatment of cancer. Consequently, there is an unmet need for the identification and generation of effective small compounds which selectively inhibit the signal transduction of the KDR/FLK-1 receptor in order to effectively and specifically suppress vasculogenesis.
Consequently, there is an unmet need for the identification and generation of effective small compounds which selectively inhibit the signal transduction of the KDR/FLK-1 receptor in order to effectively and specifically suppress vasculogenesis.
Some of the compounds of the present invention demonstrate excellent activity in biological assays and thus these compounds and related compounds are expected to be effective in treating Flk related disorders such as those driven by persistent unregulated or inappropriate angiogenesis.
The compounds described herein can be administered to a human patient per se, or in pharmaceutical compositions where they are mixed with other active ingredients, as in combination therapy, or suitable carriers or excipient(s). Techniques for formulation and administration of the compounds of the instant application may be found in xe2x80x9cRemington""s Pharmaceutical Sciences,xe2x80x9d Mack Publishing Co., Easton, Pa., latest edition.
a) Routes Of Administration
Suitable routes of administration may, for example, include oral, rectal, transmucosal, or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, intravenous, intramedullary injections, as well as intrathecal, direct intraventricular, intraperitoneal, intranasal, or intraocular injections.
Alternately, one may administer the compound in a local rather than systemic manner, for example, via injection of the compound directly into a solid tumor, often in a depot or sustained release formulation.
Furthermore, one may administer the drug in a targeted drug delivery system, for example, in a liposome coated with tumor-specific antibody. The liposomes will be targeted to and taken up selectively by the tumor.
b) Composition/Formulation
The pharmaceutical compositions of the present invention may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.
Pharmaceutical compositions for use in accordance with the present invention thus may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.
For injection, the agents of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks""s solution, Ringer""s solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained by mixing one or more solid excipient with one or more compound of the invention, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration.
For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.
For administration by inhalation, the compounds for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
The compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection maybe presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.
In addition to the formulations described previously, the compounds may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
A pharmaceutical carrier for the hydrophobic compounds of the invention is a cosolvent system comprising benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. The cosolvent system may be the VPD co-solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80(trademark), and 65% w/v polyethylene glycol 300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD:D5W) consists of VPD diluted 1:1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. Naturally, the proportions of a co-solvent system may be varied considerably without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components may be varied: for example, other low-toxicity nonpolar surfactants may be used instead of Polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.
Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide also may be employed, although usually at the cost of greater toxicity. Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.
Many of the PK modulating compounds of the invention may be provided as salts with pharmaceutically compatible counterions. Pharmaceutically compatible salts may be formed with many acids, including but not limited to hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts tend to be more soluble in aqueous or other protonic solvents than are the corresponding free base forms.
c) Effective Dosage.
Pharmaceutical compositions suitable for use in the present invention include compositions where the active ingredients are contained in an amount effective to achieve its intended purpose. More specifically, a therapeutically effective amount means an amount of compound effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.
For any compound used in the methods of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. For example, a dose can be formulated in animal models to achieve a circulating concentration range that includes the IC50 as determined in cell culture (i.e., the concentration of the test compound which achieves a half-maximal inhibition of the PK activity). Such information can be used to more accurately determine useful doses in humans.
Toxicity and therapeutic efficacy of the compounds described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between LD50 and ED50. Compounds which exhibit high therapeutic indices are preferred. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in human. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient""s condition. (See e.g., Fingl et al., 1975, in xe2x80x9cThe Pharmacological Basis of Therapeuticsxe2x80x9d, Ch. 1 p.1).
Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the kinase modulating effects, or minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vitro data; e.g., the concentration necessary to achieve 50-90% inhibition of the kinase using the assays described herein. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations.
Dosage intervals can also be determined using MEC value. Compounds should be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%.
In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.
The amount of composition administered will, of course, be dependent on the subject being treated, on the subject""s weight, the severity of the affliction, the manner of administration and the judgment of the prescribing physician.
d) Packaging
The compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the polynucleotide for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition. Suitable conditions indicated on the label may include treatment of a tumor, inhibition of angiogenesis, treatment of fibrosis, diabetes, and the like.
The compounds of the present invention were tested for their ability to inhibit most of protein kinase activity. The biological assays and results of these inhibition studies are reported herein. The methods used to measure modulation of protein kinase function are similar to those described in International Publication No. WO 98/07695, published Mar. 26, 1998, by Tang et al., and entitled xe2x80x9cIndolinone Combinatorial Libraries and Related Products and Methods for the Treatment of Disease,xe2x80x9d with respect to the high throughput aspect of the method. The WO 98/07695 publication is incorporated herein by reference in its entirety, including any drawings.
Methods of preparing pharmaceutical formulations of the compounds, methods of determining the amounts of compounds to be administered to a patient, and modes of administering compounds to an organism are disclosed in the WO 98/07695 publication, and International patent publication number WO 96/22976, by Buzzetti et al., and entitled xe2x80x9cHydrosoluble 3-Arylidene-2-Oxindole Derivatives as Tyrosine Kinase Inhibitors,xe2x80x9d published Aug. 1, 1996, both of which are incorporated herein by reference in their entirety, including any drawings. Those skilled in the art will appreciate that such descriptions are applicable to the present invention and can be easily adapted to it.